Patent application title: METHODS OF MODULATING STOMATA CONDUCTANCE AND PLANT EXPRESSION CONSTRUCTS FOR EXECUTING SAME
Inventors:
David Granot (Jerusalem, IL)
Gilor Kelly (Beit-Elazari, IL)
Menachem Moshelion (Rechovot, IL)
IPC8 Class: AC12N1582FI
USPC Class:
800289
Class name: Multicellular living organisms and unmodified parts thereof and related processes method of introducing a polynucleotide molecule into or rearrangement of genetic material within a plant or plant part the polynucleotide confers resistance to heat or cold (e.g., chilling, etc.)
Publication date: 2014-11-20
Patent application number: 20140345000
Abstract:
Plant expression construct are provided. According to an embodiment, the
plant expression construct comprises a nucleic acid sequence encoding a
hexokinase under a transcriptional control of a guard cell-specific
cis-acting regulatory element. Also provided are methods of using the
constructs and transgenic plants, plant cells and plant parts expressing
same.Claims:
1. A plant expression construct comprising a nucleic acid sequence
encoding a hexokinase under a transcriptional control of a guard
cell-specific cis-acting regulatory element.
2. A plant expression construct comprising a nucleic acid sequence encoding a nucleic acid agent for silencing expression of a hexokinase, wherein expression of said nucleic acid agent is under a transcriptional control of a guard cell-specific cis-acting regulatory element.
3-4. (canceled)
5. The plant expression construct of claim 1, wherein said guard cell-specific cis-acting regulatory element is a guard-cell specific promoter.
6. The plant expression construct of claim 5, wherein said guard-cell specific promoter is KST1 promoter.
7. A method of regulating plant stomata conductance, the method comprising modulating in the plant the level and/or activity of a hexokinase in a guard cell specific manner, thereby regulating plant conductance.
8. The method of claim 7, wherein said modulating is upregulating.
9. The method of claim 8, wherein said upregulating is effected by introducing the nucleic acid construct comprising a nucleic acid sequence encoding a hexokinase under a transcriptional control of a guard cell-specific cis-acting regulatory element into the plant.
10. The method of claim 7, wherein said modulating is downregulating.
11. The method of claim 9, wherein said downregulating is effected by introducing into the plant a nucleic acid silencing agent under a transcriptional control of a guard cell-specific cis-acting regulatory element.
12. A method of decreasing plant stomata conductance, the method comprising introducing into a cell of a plant the nucleic acid construct of claim 1, thereby decreasing the stomata conductance of the plant.
13. A method of increasing water use efficiency of a plant, the method comprising introducing into a cell of the plant the nucleic acid construct of claim 1, thereby increasing water use efficiency of the plant.
14. A method of increasing tolerance of a plant to drought, salinity or temperature stress, the method comprising introducing into a cell of the plant the nucleic acid construct of claim 1, thereby increasing tolerance of the plant to drought, salinity or temperature stress.
15. A method of increasing biomass, vigor or yield of a plant, the method comprising introducing into a cell of the plant the nucleic acid construct of claim 1, thereby increasing the biomass, vigor or yield of the plant.
16. A method of increasing tolerance of a plant to biotic stress, the method comprising introducing into a cell of the plant the nucleic acid construct of claim 1, thereby increasing tolerance of the plant to biotic stress.
17. A transgenic plant or a part thereof comprising the plant expression construct of claim 1.
18-19. (canceled)
20. The part of the transgenic plant of claim 17, being a seed.
21. The part of the transgenic plant of claim 17, being a leaf.
22. The part of the transgenic plant of claim 17, wherein said seed is a hybrid seed.
23. The method of claim 12, further comprising growing the plant under water deficient conditions or under salinity.
24. (canceled)
Description:
[0001] This application claims the benefit of priority under 35 USC 119(e)
of U.S. Provisional Patent Application No. 61/569,251 filed Dec. 11,
2011, the contents of which are incorporated herein by reference in their
entirety.
FIELD AND BACKGROUND OF THE INVENTION
[0002] The present invention, in some embodiments thereof, relates to methods of modulating stomata conductance and plant expression constructs for executing same.
[0003] Stomata are dynamic pores in the impermeable protective cuticle that coats the aerial parts of land plants, which evolved primarily to save water. Stomata, which are comprised of two guard cells and the pore they circumscribe, open at dawn to allow the entry of atmospheric carbon dioxide (CO2) for photosynthesis, at the cost of extensive transpirational water loss. The stomata close when carbon fixation and utilization are less efficient, in order to reduce the loss of water via transpiration (Assmann, 1993). At the mechanistic level, stomata open in response to increases in the osmolarity of the guard cells. These increases in osmolarity are followed by the movement of water into the guard cells, which increases their volume and opens the stomata (Taiz and Zeiger, 1998). Stomatal closure occurs in the opposite manner; as the osmolarity of guard cells is reduced, their volume decreases.
[0004] Water scarcity is a serious problem that will be exacerbated by global climate change. Abiotic stresses, especially drought and increased salinity, are primary causes of crop loss worldwide. Moreover, agriculture currently uses over 70% (86% in developing countries) of available freshwater. One of the approaches that may be adopted to save water in agriculture is the development of plants that use less water yet maintain high yields in conditions of water scarcity. As plants lose over 95% of their water via transpiration through stomata, the engineering of stomatal activity is a promising approach to reduce the water requirement of crops and to enhance productivity under stress conditions.
[0005] Cominelli et al. Transcription. 2010 July-August; 1(1): 41-45 reviews recent developments in the identification of transcription regulators controlling stomatal movements and involved in stomatal closure.
[0006] Additional background art includes
[0007] U.S. Pat. No. 7,423,203 teaches a method of increasing plant yield by expressing fungal hexokinase under a seed-specific promoter.
[0008] U.S. Patent Application 20090265812 teaches a method of increasing plant tolerance to high temperature stress by expressing hexokinase under a pollen specific promoter.
SUMMARY OF THE INVENTION
[0009] According to an aspect of some embodiments of the present invention there is provided a plant expression construct comprising a nucleic acid sequence encoding a hexokinase under a transcriptional control of a guard cell-specific cis-acting regulatory element.
[0010] According to an aspect of some embodiments of the present invention there is provided a plant expression construct comprising a nucleic acid sequence encoding a nucleic acid agent for silencing expression of a hexokinase, wherein expression of the nucleic acid agent is under a transcriptional control of a guard cell-specific cis-acting regulatory element.
[0011] According to some embodiments of the invention, the guard cell-specific cis-acting regulatory element is inducible.
[0012] According to some embodiments of the invention, the guard cell-specific cis-acting regulatory element is constitutive.
[0013] According to some embodiments of the invention, the guard cell-specific cis-acting regulatory element is a guard-cell specific promoter.
[0014] According to some embodiments of the invention, the guard-cell specific promoter is KST1 promoter.
[0015] According to an aspect of some embodiments of the present invention there is provided a method of regulating plant stomata conductance, the method comprising modulating in the plant the level and/or activity of a hexokinase in a guard cell specific manner, thereby regulating plant conductance.
[0016] According to some embodiments of the invention, the modulating is upregulating.
[0017] According to some embodiments of the invention, the upregulating is effected by introducing the nucleic acid construct of claim 1 into the plant.
[0018] According to some embodiments of the invention, the modulating is downregulating.
[0019] According to some embodiments of the invention, the downregulating is effected by introducing into the plant a nucleic acid silencing agent under a transcriptional control of a guard cell-specific cis-acting regulatory element.
[0020] According to an aspect of some embodiments of the present invention there is provided a method of decreasing plant stomata conductance, the method comprising introducing into a cell of a plant the nucleic acid construct, thereby decreasing the stomata conductance of the plant.
[0021] According to an aspect of some embodiments of the present invention there is provided a method of increasing water use efficiency of a plant, the method comprising introducing into a cell of the plant the nucleic acid construct, thereby increasing water use efficiency of the plant.
[0022] According to an aspect of some embodiments of the present invention there is provided a method of increasing tolerance of a plant to drought, salinity or temperature stress, the method comprising introducing into a cell of the plant the nucleic acid construct, thereby increasing tolerance of the plant to drought, salinity or temperature stress.
[0023] According to an aspect of some embodiments of the present invention there is provided a method of increasing biomass, vigor or yield of a plant, the method comprising introducing into a cell of the plant the nucleic acid construct, thereby increasing the biomass, vigor or yield of the plant.
[0024] According to an aspect of some embodiments of the present invention there is provided a method of increasing tolerance of a plant to biotic stress, the method comprising introducing into a cell of the plant the nucleic acid construct, thereby increasing tolerance of the plant to biotic stress.
[0025] According to an aspect of some embodiments of the present invention there is provided a transgenic plant or a part thereof comprising the plant expression construct.
[0026] According to an aspect of some embodiments of the present invention there is provided an isolated plant cell or a plant cell culture comprising the plant expression construct.
[0027] According to some embodiments of the invention, the part of the transgenic plant is a seed.
[0028] According to some embodiments of the invention, the part of the transgenic plant is a leaf.
[0029] According to some embodiments of the invention, the seed is a hybrid seed.
[0030] According to some embodiments of the invention, the method further comprises growing the plant under water deficient conditions.
[0031] According to some embodiments of the invention, the method further comprises growing the plant under salinity.
[0032] Unless otherwise defined, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.
BRIEF DESCRIPTION OF THE DRAWINGS
[0033] Some embodiments of the invention are herein described, by way of example only, with reference to the accompanying drawings. With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of embodiments of the invention. In this regard, the description taken with the drawings makes apparent to those skilled in the art how embodiments of the invention may be practiced.
[0034] In the drawings:
[0035] FIGS. 1A-C are graphs showing that sucrose stimulates stomatal closure via hexokinase. FIG. 1A--Representative light microscopy images of stomata taken from epidermal imprints 3 h after treatment with 100 mM sorbitol or 100 mM sucrose (white bar=20 μm). B, Stomatal response to sucrose in wild-type (WT) and AtHXK1-expressing plants (HK4) was assayed with intact leaves immersed for 3 h in artificial apoplastic sap (Wilkinson and Davies, 1997) containing 100 mM sorbitol (as an osmotic control), 100 mM Suc or 100 mM sucrose together with 20 mM of the hexokinase inhibitor N-acetyl-glucoseamine (NAG). Epidermal imprints were then taken and stomatal aperture was measured. C, The stomatal responses of WT plants to the different sugar combinations were assayed as described in (FIG. 1B), with 200 mM mannitol serving as an additional osmotic control. The data shown in FIGS. 1B, C are means of 300 stomata from six independent biological repeats±SE. Different letters indicate a significant difference (t test, P<0.05).
[0036] FIGS. 2A-D show that elevated expression of hexokinase enhances stomatal closure and decreases transpiration. Stomatal aperture (FIG. 2A) and stomatal conductance (FIG. 2B) were determined for control (WT) and transgenic plants expressing different levels of AtHXK1 (HK38>HK4>HK37) (Dai et al., 1999). Aperture data are means of 200 stomata from four independent repeats±SE. Stomatal conductance data are means of six independent repeats±SE. Different letters indicate a significant difference (t test, P<0.05). FIG. 2C--The rate of transpiration normalized to the total leaf area was monitored simultaneously and continuously throughout the day and the data are given as the means±SE for each 10th sampling point (n=6). FIG. 2D-A negative correlation was observed between whole-plant relative daily transpiration and relative hexokinase-phosphorylation activity. The transpiration data were normalized to the total leaf area and the amount of water taken up by the neighboring submerged fixed-size wick each day, which was set to 100%. WT hexokinase activity was set to 100%.
[0037] FIGS. 3A-E show that AtHXK1 reduces transpiration primarily when expressed in leaves. Reciprocal grafting (FIG. 3A) and triple-grafting (FIG. 3D) procedures were performed at the seedling stage and plants were photographed and used for transpiration measurements about 4 weeks after grafting. The yellow arrows and brackets indicate the location of the grafts. FIG. 3B-Whole-plant relative daily transpiration of reciprocal-grafted plants. Data were normalized to the total leaf area and the amount of water taken up by the neighboring submerged fixed-size wick each day, which was set to 100%. Data are given as means of four independent repeats±SE. Different letters indicate a significant difference (t test, P<0.05). FIG. 3C--Transpiration rate normalized to the total leaf area of reciprocal-grafted plants was monitored simultaneously and continuously throughout the day. The data are given as the means±SE for each 10th sampling point (n=4). FIG. 3E--Relative daily transpiration of whole triple-grafted plants calculated as in (FIG. 3B).
[0038] FIGS. 4A-B are graphs showing that suppression of HXK inhibits stomatal closure in response to Suc. FIG. 4A--Quantitative measurements of the real-time expression of tomato LeHXK1-3 genes in wild-type tomato (WT) and in two independent tomato lines with antisense suppression of HXK, αHK1 and αHK2. Data are means of three independent biological repeats±SE. Asterisks denote significant differences relative to the WT (t test, P<0.05). FIG. 4B-Stomatal response to Suc in WT, two antisense (αHK1 and αHK2) and AtHXK1-expressing (HK4) lines was assayed in intact leaves that were immersed in artificial apoplastic sap (Wilkinson and Davies, 1997) containing 100 mM Suc for 3 h. Data are given as means of 400 stomata from eight independent biological repeats±SE. Different letters indicate a significant difference (t test, P<0.05).
[0039] FIG. 5 is a graph showing that glucose (Glc) and sugars that can be phosphorylated, but not metabolized, stimulate stomatal closure. Stomatal responses to different sugars were assayed in intact leaves of wild-type plants. The leaves were immersed for 3 h in artificial apoplastic sap (Wilkinson and Davies, 1997) containing mannitol (as an osmotic control), Glc, 2-deoxyglucose (2-dG) or mannose. Epidermal imprints were then taken and stomatal aperture was measured. Data are given as means of 400 stomata from eight independent biological repeats±SE. Different letters indicate a significant difference (t test, P<0.05).
[0040] FIGS. 6A-F show that Suc stimulates ABA-dependent NO production in guard cells that is mediated by HXK. FIGS. 6A-B--Nitric oxide (NO) levels were monitored in guard cells from epidermal peels of wild-type (WT) and AtHXK1-expressing (HK4) plants using the fluorescent NO indicator dye DAF-2DA. Relative fluorescence levels of guard cells (white bars) and stomatal apertures (black bars) were determined after 30 min of treatment with MES buffer (control) or MES containing either 100 mM Suc or 100 mM sorbitol as an osmotic control. Representative fluorescence images are shown above the fluorescence columns (bar=10 μm). Data are given as means±SE of 90 stomata (FIG. 6A) or 60 stomata (FIG. 6B) for each treatment with three to four independent biological repeats of each treatment. FIG. 6C--Relative fluorescence levels of WT guard cells were determined after 30 min of treatment with MES buffer (control), MES containing 20 mM of the hexokinase inhibitor N-acetyl-glucoseamine (NAG), or 100 mM Suc with or without 20 mM NAG. Representative fluorescence images are shown above the fluorescence columns (bar=10 μm). Data are given as means of 60 stomata from three independent biological repeats per treatment±SE. FIG. 6D--Confocal images of NO production in guard cells of epidermal peels treated with 20 mM NAG only (left), 30 min after the addition of 100 mM Suc (middle) and 30 min after the NAG was washed out with 100 mM Suc (right). The assay was conducted as the same epidermal strip was being photographed (bar=20 μm). FIG. 6E--Relative fluorescence levels of guard cells from an epidermal strip treated as in (FIG. 6D). Data are given as means of 40-60 stomata±SE. FIG. 6F--Confocal images of NO production in guard cells of epidermal peels of Sitiens (ABA-deficient mutants) after 30 min of treatment with MES buffer containing either 100 mM Suc (left) or 100 μM ABA (right); bar=10 μm. Different lower-case letters in (FIGS. 6A-C, E) indicate a significant difference among the treatments with respect to the fluorescence data and different upper-case letters in (FIG. 6A) indicate a significant difference among the treatments with respect to the stomatal aperture data (t test, P<0.05).
[0041] FIGS. 7A-E show that GFP expression under the control of the KST1 promoter is specific to guard cells. FIG. 7A--Confocal images of wild-type (WT) (panels 1-4) and transgenic tomato leaves (panels 5-8) of plants with guard-cell specific expression of GFP (designated GCGFP) under the control of the KST1 promoter. Panels 1 and 5 are images of GFP fluorescence (stained green), panels 2 and 6 are chlorophyll autofluorescence (stained magenta), panels 3 and 7 are white light images and panels 4 and 8 are merged images. B-E, Confocal images of WT (left) and transgenic Arabidopsis GCGFP plants (right). Images were taken from leaves (FIGS. 7B and C, bars=50 μm and 5 μm, respectively), hypocotyls (FIG. 7D, bar=100 μm) and roots (FIG. 7E, bar=50 μm). All panels are merged images of white light, chlorophyll autofluorescence (magenta) and GFP fluorescence (green).
[0042] FIGS. 8A-F show that guard cell-specific expression of AtHXK1 induces stomatal closure and reduces transpiration of tomato and Arabidopsis plants. FIG. 8A--Representative images of wild-type (WT) and two independent transgenic tomato lines expressing AtHXK1 specifically in guard cells (GCHXK7 and 12). FIGS. 8B and C--Stomatal conductance (gs,) and whole-plant relative daily transpiration of WT and two independent transgenic tomato lines (GCHXK7 and 12). Stomatal conductance data are given as means of four independent repeats±SE. Transpiration data were normalized to the total leaf area and the amount of water taken up by the neighboring submerged fixed-size wick each day, which was set to 100%. Data from three consecutive days are presented. Data for each day are given as means of four independent repeats±SE. FIG. 8D--Representative images of WT Arabidopsis (Col. ecotype) and two independent transgenic lines expressing AtHXK1 specifically in guard cells (GCHXK1 and 2). FIGS. 8E and F--Stomatal conductance and transpiration measurements of WT, two independent transgenic Arabidopsis lines, GCHXK1 and GCHXK2 (Col. ecotype), and of the gin 2-1 (AtHXK1 null mutant, Ler. ecotype). Arrows indicate increased or decreased conductance and transpiration relative to the WT. Data are given as means (±SE) of 8 and 12 independent repeats for the GCHXK and gin2-1 lines, respectively. Asterisks denote significant differences relative to the WT (t test, P<0.05).
[0043] FIG. 9 shows that GFP expression under the control of the FBPase promoter is specific to mesophyll cells. Confocal images of transgenic tomato and Arabidopsis leaves of plants with mesophyll specific expression of GFP (designated MCGFP) under the control of the FBPase promoter. Images are merge of GFP fluorescence (stained green) and white light images (bar=100 μm). Fluorescence is specific to mesophyll cells.
[0044] FIGS. 10A-D are graphs showing that elevated expression of hexokinase in guard cells reduces transpiration while photosynthesis remains unchanged, thus improving instantaneous water use efficiency. Gas exchange analysis of GCHXK and WT plants was assayed using a Li-6400 portable gas-exchange system (LI-COR), stomatal conductance (FIG. 10A), transpiration (FIG. 10B), photosynthesis (FIG. 10C) and instantaneous water use efficiency (IVVUE, FIG. 10D) were measured and calculated under favorable growth conditions. Data are mean±SE (n=10 for WT and n=20 for 10 different transgenic lines, two measurements each). Star denotes significant difference (t test, P<0.05).
[0045] FIGS. 11A-C show that elevated expression of hexokinase in guard cells reduces whole plant transpiration and increases water use efficiency. FIGS. 11A-B--Whole plant relative daily transpiration (RDT) was analyzed using the large-scale lysimeter system as described in Example 1. WT and two GCHXK transgenic lines (GCHXK7, GCHXK12) were put on scales. Transpiration and total plant weight were documented every 3 minutes during the experiment in which plants were grown under normal conditions for 10 days, than subjected to drought stress for 3 days, followed by recovery irrigation process for additional 7 days. Data were normalized to the total plant weight and the amount taken up by the neighboring submerged fixed-size wick each day, which was set to 100% and served as a reference for the temporal variations in the potential transpiration. FIG. 11A--Day by day Relative daily transpiration during the whole experiment. Data are means of four independent repeats±SEM FIG. 11B--Relative daily transpiration of selected days in each treatment. Data are means of four independent repeats±SEM; Star denotes significant difference (t test, P<0.05). FIG. 11C--Water use efficiency was calculated by the ratio between plant weight accumulation and plant water loss, each day per each plant. Data are means of four independent repeats±SEM; Star denotes significant difference (t test, P<0.05). (A-magnified) RDT of WT and GCHXK plants during the shift from normal irrigation (day 10) to drought conditions (day 11). Red and green arrows indicate RDT decline (represented by slope) of WT and GCHXK respectively after plants were exposed to drought.
[0046] FIGS. 12A-F show that elevated expression of hexokinase in guard cells reduces transpiration rate and stomatal conductance throughout the day, while displaying normal growth. Whole plant relative transpiration rate (FIG. 12A) and stomatal conductance (gs, FIG. 12B) were analyzed using the large-scale lysimeter system as described in methods. WT and two GCHXK transgenic lines were put on scales. Transpiration rate, gs, light intensity (FIG. 12E), vapor pressure deficit (VPD, FIG. 12F) were simultaneously documented every 3 minutes during the experiment in which plants were grown under normal conditions. Data for FIGS. 12A and B were normalized to the total leaf area and the amount taken up by the neighboring submerged fixed-size wick each day, which was set to 100% and served as a reference for the temporal variations in the potential transpiration. FIG. 12C--Total plant leaf area, FIG. 12D--Total plant weight.
[0047] FIG. 13 show transpiration rate of WT and GCHXK plants under drought conditions. Whole plant transpiration rate was analyzed using the large-scale lysimeter system as described in Example 1. WT (blue) and GCHXK transgenic lines (green) were put on scales. Transpiration rates were documented for 9 days after exposing the plants to gradually increased--drought conditions, by fully stopping the irrigation. The rate of transpiration normalized to the total leaf area was monitored simultaneously and continuously throughout the day and the data are given as the means±SE for each sampling point. Data were normalized to the total leaf area and the amount taken up by the neighboring submerged fixed-size wick each day, which was set to 100% and served as a reference for the temporal variations in the potential transpiration. Star denotes the day in which transpiration transition between WT and GCHXK had occurred.
[0048] FIGS. 14A-B show the yield production of transgenic plants expressing hexokinase specifically in guard cells. FIG. 14A--Number of fruits collected from WT and GCHXK plants (4 independent lines). FIG. 14B--Representative images of wild-type (WT) and transgenic tomato plant expressing AtHXK1 specifically in guard cells (GCHXK7).
[0049] FIGS. 15A-C show the yield production of transgenic plants expressing hexokinase specifically in guard cells, under limited water-supply conditions. FIG. 15A--Plants were grown under controlled commercial greenhouse conditions, following expert instructions with regard to growing procedures (Soil system, irrigation, fertilization etc.). Seedlings were planted in a mixed up order threw out the entire planting-row and the same order was kept in each row. Each row was irrigated differentially; either fully (100%) or partially (75%, 50% and 25% irrigation regimes). Since the initial fruit breaker stage, fruits were collected, counted and weighted for each individual plant for 4 weeks time. Cumulative fruit weight (FIG. 15B) and fruit number (FIG. 15C) of WT (blue) and GCHXK (green) plants were than averaged for each irrigation regime. Blue and green arrows indicates decreased fruit weight of WT and GCHXK plants respectively when shifting from 75% to 50% irrigation.
[0050] FIGS. 16A-F show that guard cell-specific expression of AtHXK1 induces stomatal closure, reduces transpiration and increases leaf temperature without lowering photosynthesis or mesophyll conductance for CO2, thus enhances water use efficiency of Arabidopsis plants. Stomatal conductance (FIG. 16A), transpiration (FIG. 16B), photosynthesis (FIG. 16C) and mesophyll conductance for CO2 (gm, FIG. 16D) measurements of WT and transgenic Arabidopsis plants expressing AtHXK1 specifically in guard cells (GCHXK). FIG. 16E--instantaneous water use efficiency (IWUE) of WT and GCHXK plants. FIG. 16F--Leaf temperatures (warmer leaves--stomatal closure) of WT and GCHXK plants were determined using ThermaCam researcher pro 2.10 software. Data points are the means±SE from 6 biological repeats in FIGS. 16A-E and of 12 biological repeats in FIG. 16F. An asterisk denotes a significant difference relative to the wild type (t test, P<0.05).
[0051] FIGS. 17A-B are schematic maps of binary vector pGreen0029 containing KST1 promoter, AtHXK1 cDNA (FIG. 17A) or GFP (FIG. 17B) and a terminator: Vector also contains nos-Kan and neomycin phosphotransferase II (NptH) genes as selectable markers for bacteria and plant transformation.
DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION
[0052] The present invention, in some embodiments thereof, relates to methods of modulating stomata conductance and plant expression constructs for executing same.
[0053] Before explaining at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details set forth in the following description or exemplified by the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways.
[0054] Water is the major factor limiting the growth and development of many land plants. Stomata, composed of two guard cells, are the chief gates controlling plants' water loss. Many environmental and physiological stimuli control stomatal opening, but they all function through the regulation of guard-cell osmolarity. Increased guard-cell osmolarity leads to the opening of the stomata and decreased osmolarity causes the stomata to close. The prevailing paradigm is that sucrose acts as an osmoticum in the guard cells, thereby contributing to the opening of the stomata.
[0055] While conceiving the present invention, the present inventors have found that contrary to the prevailing paradigm, sucrose closes stomata via a non-osmotic mechanism (see Example 2). Furthermore, the guard cells' response to sucrose is dependent on the sugar-sensing enzyme hexokinase (HXK), which triggers the abscisic acid-signaling pathway within the guard cells, leading to stomatal closure.
[0056] Thus, while reducing the present invention to practice, the present inventors have found that modulation of hexokinase activity or expression correlates with stomatal aperture.
[0057] As is illustrated herein below and in the Examples section which follows, the present inventors have overexpressed HXK in the stomata of tomato plants (in a guard-cell specific manner). Surprisingly, while photosynthesis remained unchanged (FIG. 10C), stomatal conductance (indicating stomatal aperture, FIG. 10B) and transpiration (FIG. 10A) were reduced. Similar results were obtained while monitoring the same parameters all day long (FIGS. 12A-D). Importantly, by measuring total plant leaf area and weight (FIGS. 12C and 12D respectively), the present inventors discovered that even though plants have consumed less water (FIG. 12A), growth was not impaired, and was even improved. Saving water without affecting plant growth improves whole plant water use efficiency. Elevated expression of hexokinase in guard cells improves yield production (FIGS. 14A-B) even under limited water supply (FIGS. 15A-C). Similar results were observed in Arabidopsis. These results demonstrate that the same transgenic insertion of hexokinase under guard-cell specific promoter used in the case of Tomato (Solanaceae family) is universally applicable while affecting stomata and increases water use efficiency in the case of Arabidopsis (Brassicaceae family) as well, and that this technique could be implemented in other species as well.
[0058] Unlike previous studies, which relied on correlations between sucrose content and stomatal aperture, this study took a functional approach to the examination of the effects of sucrose and its cleavage products on stomatal behavior. It is now proven that sucrose stimulates a guard cell-specific response that is mediated by HXK and ABA and leads to stomatal closure. Without being bound to theory it is suggested that this response presents a natural feedback mechanism aimed at reducing transpiration and conserving water under excess of photosynthesis, thus coordinating between photosynthesis and transpiration.
[0059] Thus, according to an aspect of the invention there is provided a method of regulating plant stomata conductance, the method comprising modulating in the plant the level and/or activity of a hexokinase in a guard cell specific manner, thereby regulating stomata conductance and plant transpiration.
[0060] As used herein the phrase "stomata conductance" refers to gaseous exchange through the stomata pore complex. Stomata conductance is regulated by stomata aperture. Stomatal conductance affects plant transpiration and therefore the present methodology according to this aspect of the invention also regulated plant transpiration.
[0061] As used herein the phrase "regulating plant stomata conductance" refers to increase or decrease in stomata conductance. The increase or decrease may be by at least 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or more say 90% or 100% (e.g., 20-40%).
[0062] As used herein the term "hexokinase", abbreviated as HXK, and referred to herein as "the transgene" or "the polypeptide", refers to the enzyme that typically phosphorylates hexoses (six-carbon sugars), forming hexose phosphate and having the E.C. Number 2.7.1.1. HXK as used herein also refers to hexokinase-like (HKL) protein that binds hexose and transmits a signal independent of its kinase (hexose phosphorylation) activity.
[0063] Hexokinases according to the present teachings may be about 100 kD in size as of most multicellular organisms (e.g., mammalian and plants). They consist of two halves (N and C terminal), which share much sequence homology. This suggests an evolutionary origin by duplication and fusion of a 50 kD ancestral hexokinase similar.
[0064] The hexokinase may be naturally occurring or may comprise/consist of a synthetic sequence (i.e., man-made) as long as it retains a hexokinase activity.
[0065] Due to their high conservation level, the hexokinase of the present invention can be of a plant- or an animal origin. According to a specific embodiment, the hexokinase is a plant hexokinase.
[0066] The hexokinases can be categorized according to their cellular localization. Thus, the HXKs may be associated with the mitochondria, associated with or within plastids or present in the cytosol. To date, all of the HXKs examined in eudicots have been found to have either a plastidic signal peptide (type A) or an N-terminal membrane anchor domain (type B), however, cytosolic hexokinases are also contemplated for use according to the present teachings. According to a specific embodiment, the hexokinase is a type B (mitochondrial associated) HXK.
[0067] As used herein "a hexokinase activity" refers to the ability of the enzyme to regulate stomata conductance. The enzyme may bind hexose and stimulate the abscisic acid (ABA) pathway which controls stomata conductance. The activity may be kinase independent.
[0068] Non limiting examples of hexokinases which are contemplated according to the present teachings are provided in Table 1 herein below.
TABLE-US-00001 TABLE 1 Hexokinase genes and their physiological function. Accession Type/ no. Intracellular Physiological Species Gene (SEQ ID NO:) localization function References Eudicots Arabidopsis AtHXK1 AT4G29130 Type B Glc sensing (Jang et al., 1997; Dai et al., thaliana (SEQ ID NOs: M, N PCD 1999; Yanagisawa et al., 11 and 12) Mediates sugar and 2003; Moore et al., 2003; hormonal interactions Leon and Sheen, 2003; Kim Growth and et al., 2006; Pourtau et al., development 2006; Cho et al., 2006a; Photosynthetic gene Rolland et al., 2006; Chen, repression 2007; Aki et al., 2007; Transpiration Balasubramanian et al., Actin filament 2007, 2008; Sarowar et al., reorganization 2008; Karve et al., 2008; Ju Oxidative stress et al., 2009; Karve et al., response 2010; Kushwah et al., 2011; Pathogen resistance Kelly et al., 2012) Directional root growth Leaf senescence AtHXK2 AT2G19860 Type B Glc sensing (Jang et al., 1997; Kim et (SEQ ID NOs: M PCD al., 2006; Karve et al., 13 and 14) Photosynthetic gene 2008) repression AtHXK3 AT1G47840 Type A Glc sensing (Claeyssen and Rivoal, (SEQ ID NOs: P Abiotic stress 2007; Karve et al., 2008; 15 and 16) response Zhang et al., 2010) AtHKL1 AT1G50460 Type B Growth (Claeyssen and Rivoal, (SEQ ID NOs: M Root hair 2007; Karve et al., 2008; 17 and 18) development Karve and Moore, 2009; Mediates Glc- Karve et al., 2012) ethylene crosstalk Abiotic stress response AtHKL2 AT3G20040 Type B (Karve et al., 2008) (SEQ ID NOs: M 19 and 20) AtHKL3 AT4G37840 Type B Abiotic stress (Claeyssen and Rivoal, (SEQ ID NOs: M response 2007; Karve et al., 2008) 21 and 22) Tomato SlHXK1 AJ401153 Type B (Damari-Weissler et al., (Solanum (SEQ ID NOs: M 2006) lycopersicum) 23 and 24) SlHXK2 AF208543 Type B (Menu et al., 2001; Damari- (SEQ ID NOs: M Weissler et al., 2006) 25 and 26) SlHXK3 DQ056861 Type B (Kandel-Kfir et al., 2006) (SEQ ID NOs: M 27 and 28) SlHXK4 DQ056862 Type A (Kandel-Kfir et al., 2006) (SEQ ID NOs: P 29 and 30) Solanum ScHK2 DQ177440 ND (Claeyssen et al., 2006) chacoense (SEQ ID NOs: 31 and 32) Potato StHXK1 X94302 (SEQ ND Glc sensing (Veramendi et al., 1999; (Solanum ID NOs: 33 Leaves starch content Veramendi et al., 2002) tuberosum) and 34) StHXK2 AF106068 ND Glc sensing (Veramendi et al., 2002) (SEQ ID NOs: 35 and 36) Tobacco NtHXK2 AY553215 Type A (Giese et al., 2005) (Nicotiana (SEQ ID NOs: P tabacum/ 37 and 38) benthamiana) NbHXK1 AY286011 Type B Plant growth (Kim et al., 2006; Sarowar (SEQ ID NOs: M PCD et al., 2008) 39 and 40) Oxidative-stress resistance Sunflower HaHXK1 DQ835563 ND Seed development (Troncoso-Ponce et al., (Helianthus (SEQ ID NOs: 2011) annuus) 41 and 42) Populus PtHXK1 XP_002325031 Type B Glc sensing (Karve et al., 2010) trichocarpa (SEQ ID NOs: M 43 and 44) Grape VvHXK1 JN118544 ND (Yu et al., 2012) (Vitis vinifera VvHXK2 JN118545 ND (Yu et al., 2012) L. cv. Cabernet Sauvignon) Spinach SoHXK1 AF118132 Type B (Wiese et al., 1999; (Spinacia (SEQ ID NOs: M Damari-Weissler et al., oleracea) 45 and 46) 2007) Monocots Rice OsHXK1 DQ116383 C, N (Cho et al., 2006a; Cheng et (Oryza sativa) (SEQ ID NOs: al., 2011) 47 and 48) OsHXK2 DQ116384 Type B (Cheng et al., 2011) (SEQ ID NOs: M 49 and 50) OsHXK3 DQ116385 Type B (Cheng et al., 2011) (SEQ ID NOs: M 51 and 52) OsHXK4 DQ116386 Type A (Cho et al., 2006a; Cheng et (SEQ ID NOs: P al., 2011) 53 and 54) OsHXK5 DQ116387 Type B Glc sensing (Cho et al., 2009a; Cheng et (SEQ ID NOs: M, N Photosynthetic gene al., 2011) 55 and 56) repression Shoot growth OsHXK6 DQ116388 Type B Glc sensing (Aki and Yanagisawa, (SEQ ID NOs: M, N Photosynthetic gene 2009; Cho et al., 2009a; 57 and 58) repression Cheng et al., 2011) Shoot growth OsHXK7 DQ116389 C, N (Cho et al., 2006a; Cheng et (SEQ ID NOs: al., 2011) 59 and 60) OsHXK8 DQ116390 C, N (Cheng et al., 2011) (SEQ ID NOs: 61 and 62) OsHXK9 DQ116391 Type B (Cheng et al., 2011) (SEQ ID NOs: M 63 and 64) OsHXK10 DQ116392 C Pollen germination (Xu et al., 2008; Cheng et (SEQ ID NOs: and/or al., 2011) 65 and 66) M Sorghum SbHXK3 XM_002459027 Type B No Glc sensing role (Karve et al., 2010) (Sorghum (SEQ ID NOs: M bicolor) 67 and 68) SbHXK8 XM_002454982 C (Karve et al., 2010) (SEQ ID NOs: 69 and 70) Wheat HXK AY974231 ND Controls triose (Sun et al., 2006) (Triticum (SEQ ID NOs: phosphate/phosphate aestivum) 71 and 72) translocation Lycophytes Spike moss SmHXK3 26000047 * C Glc sensing (Karve et al., 2010) (Selaginella SmHXK5 57.357.1 * C (Karve et al., 2010) mollendorffii) Bryophytes Moss PpHXK1 AY260967 Type A Filamentous type and (Olsson et al., 2003; (Physcomitrella (SEQ ID NOs: P growth Thelander et al., 2005) patens) 73 and 74) PpHXK2 XM_001784578 Type B (Nilsson et al., 2011) (SEQ ID NOs: M, P 75 and 76) PpHXK3 XM_001784282 Type B (Nilsson et al., 2011) (SEQ ID NOs: M, P 77 and 78) PpHXK4 XM_001760896 Type C (Nilsson et al., 2011) (SEQ ID NOs: C, N 79 and 80) PpHXK5 XM_001766381 Type A (Nilsson et al., 2011) (SEQ ID NOs: P 81 and 82) PpHXK6 XM_001762899 Type A (Nilsson et al., 2011) (SEQ ID NOs: P 83 and 84) PpHXK7 XM_001754096 Type B (Nilsson et al., 2011) (SEQ ID NOs: M, P 85 and 86) PpHXK8 XM_001752177 Type B (Nilsson et al., 2011) (SEQ ID NOs: M, P 87 and 88) PpHXK9 XM_001770125 Type D (Nilsson et al., 2011) (SEQ ID NOs: M 89 and 90) PpHXK10 XM_001776713 Type D (Nilsson et al., 2011) (SEQ ID NOs: M 91 and 92) PpHXK11 XM_001779426 Type D (Nilsson et al., 2011) (SEQ ID NOs: M, P 93 and 94) Type A--localized in plastid stroma. Type B--associated with the mitochondria. Type C--localized in the cytosol and nucleus. Type D--associated with the mitochondria, different from type B in sequence. M--mitochondria associated. P--plastid. N--nucleus. C--cytosol. ND--not determined. PCD--programmed cell death. Glc--glucose. *Joint Genome Institute-Selaginella moellendorffii v1.0.
[0069] As mentioned, the HXK sequence may be naturally occurring or artificially generated (e.g., codon-optimized) according to the intended use.
[0070] According to a specific embodiment, modulating the activity or expression of HXK refers to upregulating the activity or expression which results in reduction of stomatal conductance. Upregulating can be by at least 5%, 10%, 20, %, 30%, 40%, 50%, 60%, 70% 80% or more, say 90% or even 100%, as compared to hexokinase expression or activity in a similar cell of the same plant species, growth conditions and developmental stage (e.g., wild-type (WT) plant).
[0071] As mentioned, upregulation of hexokinase activity or expression in a guard-cell specific manner has a number of advantages in crop plants and vegetables farming.
[0072] Thus, the present inventors have shown that upregulation of HXK in a guard-cell specific manner decreases stomata aperture and conductance (without affecting photosynthesis), improves plant's water use efficiency, thereby increasing plant's tolerance to drought, and overall increases plants vigor, biomass or yield (under stress or optimal growth conditions). Likewise, plants expressing HXK in a guard-cell specific manner are tolerant to salinity stress. It is appreciated that Water are taken up (soaked) by plants as a result of the difference between water potential in the air and within the plants. This difference is termed vapor pressure deficit (VPD). The driving force of soaking water from the ground is the VPD. Higher VPD--the greater is the force. Yet, when the stomata are partially closed, the effect of VPD is lowered and less water is being taken up by the plant. In that case, the plant will take less salt from the ground and will be less affected. The present teachings have also an unprecedented impact on the tolerance of plants to biotic stress. Many human and plant pathogens such as bacteria and fungi, invade plants via the stomata (see for Example Kroupitski et al. Applied and Environmental Microbiology 2009 6076-6086 teaching that Salmonella enteric internalizes in leaves via open stomata). Not only does the stomata allow easy entrance, but also serve as good environment for attracting the pathogens by the accumulation of sugars near the guard cells when the stomata is open. Indeed, the present inventors have observed reduced fungi and bacteria infections in plants with high expression of HXK (not shown).
[0073] Alternatively or additionally, the present teachings can also be employed towards imparting the plant with a tolerance to temperature stress (heat or cold). For instance, plants expressing high levels of HXK in a guard cell specific manner are expected to exhibit extended heat and cold resistance with regard to fruit setting. Pollen development and germination are sensitive to heat and cold, most likely due to perturbation of sugar metabolism. It is suggested that during heat stress less sugars are being carried toward the pollen grains (and other sink tissues as well) since most of the water is transpired through the stomata. According to the present teachings, when less water is transpired through the stomata so then more water is available for sugar transport in the phloem. That may impart resistance to temperature stress (e.g., heat) thereby allowing production of viable pollen grains.
[0074] Alternatively or additionally, the present teachings can be employed towards prevention of blossom end rot (BER). BER is a visible physiological damage that affects many crops such as tomato, eggplants, pepper, melon and many more. BER happens mainly under heat and water stress. It is now suggested that under such conditions, most of the water is transpired and less water is available to carry sugars, minerals and ions toward the fruits. Accordingly, lowering transpiration may allocate more water carrying more sugars, minerals and ions toward the fruits and other sink tissues (Nikinma et al. 2012 Plant, Cell and Environment 2012 1-15). BER is determined by the percentage of fruits that exhibit visible or detectable rot (physical damage) on the fruit. BER prevention means lowering the percentage of fruits with BER.
[0075] Thus, according to an exemplary embodiment the present teachings can be used to increase biomass, vigor or yield of a plant.
[0076] As used herein the phrase "plant yield" refers to the amount (e.g., as determined by weight or size) or quantity (numbers) of tissues or organs produced per plant or per growing season. Hence increased yield could affect the economic benefit one can obtain from the plant in a certain growing area and/or growing time.
[0077] It should be noted that a plant yield can be affected by various parameters including, but not limited to, plant biomass; plant vigor; growth rate; seed yield; seed or grain quantity; seed or grain quality; oil yield; content of oil, starch and/or protein in harvested organs (e.g., seeds, fruits or vegetative parts of the plant); number of flowers (florets) per panicle (expressed as a ratio of number of filled seeds over number of primary panicles); harvest index; number of plants grown per area; number and size of harvested organs per plant and per area; number of plants per growing area (density); number of harvested organs in field; total leaf area; carbon assimilation and carbon partitioning (the distribution/allocation of carbon within the plant); resistance to shade; number of harvestable organs (e.g. seeds), seeds per pod, weight per seed; and modified architecture [such as increase stalk diameter, thickness or improvement of physical properties (e.g. elasticity)].
[0078] As used herein the phrase "seed yield" refers to the number or weight of the seeds per plant, seeds per pod, or per growing area or to the weight of a single seed, or to the oil extracted per seed. Hence seed yield can be affected by seed dimensions (e.g., length, width, perimeter, area and/or volume), number of (filled) seeds and seed filling rate and by seed oil content. Hence increase seed yield per plant could affect the economic benefit one can obtain from the plant in a certain growing area and/or growing time; and increase seed yield per growing area could be achieved by increasing seed yield per plant, and/or by increasing number of plants grown on the same given area.
[0079] The term "seed" (at times referred to as "grain" or "kernel") as used herein refers to a small embryonic plant enclosed in a covering called the seed coat (usually with some stored food), the product of the ripened ovule of gymnosperm and angiosperm plants which occurs after fertilization and some growth within the mother plant. The seed may be a hybrid seed or a homozygous line.
[0080] The phrase "oil content" as used herein refers to the amount of lipids in a given plant organ, either the seeds (seed oil content) or the vegetative portion of the plant (vegetative oil content) and is typically expressed as percentage of dry weight (10% humidity of seeds) or wet weight (for vegetative portion).
[0081] It should be noted that oil content is affected by intrinsic oil production of a tissue (e.g., seed, fruit, vegetative portion), as well as the mass or size of the oil-producing tissue per plant or per growth period.
[0082] In one embodiment, increase in oil content of the plant can be achieved by increasing the size/mass of a plant's tissue(s) which comprise oil per growth period. Thus, increased oil content of a plant can be achieved by increasing the yield, growth rate, biomass and vigor of the plant.
[0083] As used herein the phrase "plant biomass" refers to the amount (e.g., measured in grams of air-dry tissue) of a tissue produced from the plant in a growing season, which could also determine or affect the plant yield or the yield per growing area. An increase in plant biomass can be in the whole plant or in parts thereof such as aboveground (harvestable) parts, fruit biomass, vegetative biomass, roots and seeds.
[0084] As used herein the phrase "growth rate" refers to the increase in plant organ/tissue size per time (can be measured in cm2 per day).
[0085] As used herein the phrase "plant vigor" refers to the amount (measured by weight) of tissue produced by the plant in a given time. Hence increased vigor could determine or affect the plant yield or the yield per growing time or growing area. In addition, early vigor (seed and/or seedling) results in improved field stand.
[0086] It should be noted that a plant yield can be determined under stress (e.g., abiotic stress) and/or non-stress (normal) conditions. It is contemplated herein that yield, vigor or biomass of the plant expressing the HXK in a guard cell-specific manner is increased as compared to that of wild-type plant (not expressing said HXK) under stress or non-stressed conditions.
[0087] As used herein, the phrase "non-stress conditions" (or normal or optimal as referred to herein) refers to the growth conditions (e.g., water, temperature, light-dark cycles, humidity, salt concentration, fertilizer concentration in soil, nutrient supply such as nitrogen, phosphorous and/or potassium), that do not significantly go beyond the everyday climatic and other abiotic conditions that plants may encounter, and which allow optimal growth, metabolism, reproduction and/or viability of a plant at any stage in its life cycle (e.g., in a crop plant from seed to a mature plant and back to seed again). Persons skilled in the art are aware of normal soil conditions and climatic conditions for a given plant in a given geographic location. It should be noted that while the non-stress conditions may include some mild variations from the optimal conditions (which vary from one type/species of a plant to another), such variations do not cause the plant to cease growing without the capacity to resume growth.
[0088] As mentioned increased yield can be under non-stress conditions or abiotic/biotic stress conditions.
[0089] The phrase "abiotic stress" as used herein refers to any adverse effect on metabolism, growth, reproduction and/or viability of a plant. Accordingly, abiotic stress can be induced by suboptimal environmental growth conditions such as, for example, salinity, water deprivation, flooding, freezing, low or high temperature (i.e., cold or heat), heavy metal toxicity, anaerobiosis, nutrient deficiency, atmospheric pollution or UV irradiation.
[0090] The phrase "abiotic stress tolerance" as used herein refers to the ability of a plant to endure an abiotic stress without suffering a substantial alteration in metabolism, growth, productivity and/or viability.
[0091] As used herein the phrase "water use efficiency (WUE)" refers to the level of organic matter produced per unit of water consumed by the plant, i.e., the dry weight of a plant in relation to the plant's water use, e.g., the biomass produced per unit transpiration.
[0092] Similarly "biotic stress" refers to stress that occurs as a result of damage done to plants by other living organisms, such as bacteria, viruses, fungi, parasites.
[0093] Upregulation of HXK in a guard-cell specific manner can be used to remedy any of the aforementioned conditions and to improve plants performance overall. Thus, upregulation of the HXK can be effected by expressing an exogenous polynucleotide encoding HXK in the plant in a guard-cell specific manner.
[0094] The phrase "expressing within the plant an exogenous polynucleotide encoding HXK" as used herein refers to upregulating the expression level of an exogenous polynucleotide encoding an HXK polypeptide within the plant by introducing the exogenous polynucleotide into a plant cell or plant and expressing by recombinant means, as further described herein below.
[0095] As used herein "expressing" refers to expression at the mRNA and polypeptide level. It will be appreciated that for silencing the expression is at the mRNA level alone (silencing mechanisms of HXK are described further hereinbelow).
[0096] As used herein, the phrase "exogenous polynucleotide" refers to a heterologous nucleic acid sequence which may not be naturally expressed within the plant or which overexpression in the plant is desired. The exogenous polynucleotide may be introduced into the plant in a stable or transient manner, so as to produce a ribonucleic acid (RNA) molecule and/or a polypeptide molecule. It should be noted that the exogenous polynucleotide may comprise a nucleic acid sequence which is identical or partially homologous to an endogenous nucleic acid sequence of the plant.
[0097] The term "endogenous" as used herein refers to any polynucleotide or polypeptide which is present and/or naturally expressed within a plant or a cell thereof.
[0098] According to the invention, the exogenous polynucleotide of the invention comprises a nucleic acid sequence encoding a polypeptide having an amino acid sequence of a hexokinase.
[0099] According to a specific embodiment the amino acid sequence of the HXK polypeptide (encoded from the exogenous polynucleotide) is at least about, 30%, 40% or 50%, or at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more say 100% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92 and 94, as long as its hexokinase activity is maintained as described above.
[0100] Homology (e.g., percent homology, identity+similarity) can be determined using any homology comparison software, including for example, the BlastP or TBLASTN software of the National Center of Biotechnology Information (NCBI) such as by using default parameters, when starting from a polypeptide sequence; or the tBLASTX algorithm (available via the NCBI) such as by using default parameters, which compares the six-frame conceptual translation products of a nucleotide query sequence (both strands) against a protein sequence database.
[0101] According to some embodiments of the invention, the term "homology" or "homologous" refers to identity of two or more nucleic acid sequences; or identity of two or more amino acid sequences.
[0102] Homologous sequences include both orthologous and paralogous sequences. The term "paralogous" relates to gene-duplications within the genome of a species leading to paralogous genes. The term "orthologous" relates to homologous genes in different organisms due to ancestral relationship.
[0103] One option to identify orthologues in monocot plant species is by performing a reciprocal blast search. This may be done by a first blast involving blasting the sequence-of-interest against any sequence database, such as the publicly available NCBI database which may be found at: Hypertext Transfer Protocol://World Wide Web (dot) ncbi (dot) nlm (dot) nih (dot) gov. If orthologues in rice were sought, the sequence-of-interest would be blasted against, for example, the 28,469 full-length cDNA clones from Oryza sativa Nipponbare available at NCBI. The blast results may be filtered. The full-length sequences of either the filtered results or the non-filtered results are then blasted back (second blast) against the sequences of the organism from which the sequence-of-interest is derived. The results of the first and second blasts are then compared. An orthologue is identified when the sequence resulting in the highest score (best hit) in the first blast identifies in the second blast the query sequence (the original sequence-of-interest) as the best hit. Using the same rational a paralogue (homolog to a gene in the same organism) is found. In case of large sequence families, the ClustalW program may be used [Hypertext Transfer Protocol://World Wide Web (dot) ebi (dot) ac (dot) uk/Tools/clustalw2/index (dot) html], followed by a neighbor-joining tree (Hypertext Transfer Protocol://en (dot) wikipedia (dot) org/wiki/Neighbor-joining) which helps visualizing the clustering.
[0104] According to some embodiments of the invention, the exogenous polynucleotide of the invention encodes a polypeptide having an amino acid sequence at least about 30%, 40%, 50%, 60%, 70% or at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more say 100% identical to the amino acid sequence selected from the group consisting of 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92 and 94 as long as the hexokinase activity of the protein (as described above) is maintained.
[0105] As used herein the term "polynucleotide" refers to a single or double stranded nucleic acid sequence which is isolated and provided in the form of an RNA sequence, a complementary polynucleotide sequence (cDNA), a genomic polynucleotide sequence and/or a composite polynucleotide sequences (e.g., a combination of the above).
[0106] The term "isolated" refers to at least partially separated from the natural environment e.g., from a plant cell.
[0107] As used herein the phrase "complementary polynucleotide sequence" refers to a sequence, which results from reverse transcription of messenger RNA using a reverse transcriptase or any other RNA dependent DNA polymerase. Such a sequence can be subsequently amplified in vivo or in vitro using a DNA dependent DNA polymerase.
[0108] As used herein the phrase "genomic polynucleotide sequence" refers to a sequence derived (isolated) from a chromosome and thus it represents a contiguous portion of a chromosome.
[0109] As used herein the phrase "composite polynucleotide sequence" refers to a sequence, which is at least partially complementary and at least partially genomic. A composite sequence can include some exonal sequences required to encode the polypeptide of the present invention, as well as some intronic sequences interposing therebetween. The intronic sequences can be of any source, including of other genes, and typically will include conserved splicing signal sequences. Such intronic sequences may further include cis acting expression regulatory elements.
[0110] Nucleic acid sequences encoding the HXK polypeptides of the present invention may be optimized for expression. Examples of such sequence modifications include, but are not limited to, an altered G/C content to more closely approach that typically found in the plant species of interest, and the removal of codons atypically found in the plant species commonly referred to as codon optimization.
[0111] The phrase "codon optimization" refers to the selection of appropriate DNA nucleotides for use within a structural gene or fragment thereof that approaches codon usage within the plant of interest. Therefore, an optimized gene or nucleic acid sequence refers to a gene in which the nucleotide sequence of a native or naturally occurring gene has been modified in order to utilize statistically-preferred or statistically-favored codons within the plant. The nucleotide sequence typically is examined at the DNA level and the coding region optimized for expression in the plant species determined using any suitable procedure, for example as described in Sardana et al. (1996, Plant Cell Reports 15:677-681). In this method, the standard deviation of codon usage, a measure of codon usage bias, may be calculated by first finding the squared proportional deviation of usage of each codon of the native gene relative to that of highly expressed plant genes, followed by a calculation of the average squared deviation. The formula used is: 1 SDCU=n=1 N [(Xn-Yn)/Yn]2/N, where Xn refers to the frequency of usage of codon n in highly expressed plant genes, where Yn to the frequency of usage of codon n in the gene of interest and N refers to the total number of codons in the gene of interest. A Table of codon usage from highly expressed genes of dicotyledonous plants is compiled using the data of Murray et al. (1989, Nuc Acids Res. 17:477-498).
[0112] One method of optimizing the nucleic acid sequence in accordance with the preferred codon usage for a particular plant cell type is based on the direct use, without performing any extra statistical calculations, of codon optimization Tables such as those provided on-line at the Codon Usage Database through the NIAS (National Institute of Agrobiological Sciences) DNA bank in Japan (Hypertext Transfer Protocol://World Wide Web (dot) kazusa (dot) or (dot) jp/codon/). The Codon Usage Database contains codon usage tables for a number of different species, with each codon usage Table having been statistically determined based on the data present in Genbank.
[0113] By using the above Tables to determine the most preferred or most favored codons for each amino acid in a particular species (for example, rice), a naturally-occurring nucleotide sequence encoding a protein of interest can be codon optimized for that particular plant species. This is effected by replacing codons that may have a low statistical incidence in the particular species genome with corresponding codons, in regard to an amino acid, that are statistically more favored. However, one or more less-favored codons may be selected to delete existing restriction sites, to create new ones at potentially useful junctions (5' and 3' ends to add signal peptide or termination cassettes, internal sites that might be used to cut and splice segments together to produce a correct full-length sequence), or to eliminate nucleotide sequences that may negatively effect mRNA stability or expression.
[0114] The naturally-occurring encoding nucleotide sequence may already, in advance of any modification, contain a number of codons that correspond to a statistically-favored codon in a particular plant species. Therefore, codon optimization of the native nucleotide sequence may comprise determining which codons, within the native nucleotide sequence, are not statistically-favored with regards to a particular plant, and modifying these codons in accordance with a codon usage table of the particular plant to produce a codon optimized derivative. A modified nucleotide sequence may be fully or partially optimized for plant codon usage provided that the protein encoded by the modified nucleotide sequence is produced at a level higher than the protein encoded by the corresponding naturally occurring or native gene. Construction of synthetic genes by altering the codon usage is described in for example PCT Patent Application 93/07278.
[0115] The term "plant" as used herein encompasses whole plants, ancestors and progeny of the plants and plant parts (those which comprise stomata but not necessarily), including seeds, shoots, stems, roots (including tubers), and plant cells, tissues and organs. The plant may be in any form including suspension cultures, embryos, meristematic regions, callus tissue, leaves, gametophytes, sporophytes, pollen, and microspores.
[0116] According to some embodiments of the invention the plant is a dicotyledonous plant.
[0117] According to some embodiments of the invention the plant is a monocotyledonous plant.
[0118] Plants that are particularly useful in the methods of the invention include all plants which belong to the superfamily Viridiplantae, in particular monocotyledonous and dicotyledonous plants including a fodder or forage legume, ornamental plant, food crop, tree, or shrub selected from the list comprising Acacia spp., Acer spp., Actinidia spp., Aesculus spp., Agathis australis, Albizia amara, Alsophila tricolor, Andropogon spp., Arachis spp, Areca catechu, Astelia fragrans, Astragalus cicer, Baikiaea plurijuga, Betula spp., Brassica spp., Bruguiera gymnorrhiza, Burkea africana, Butea frondosa, Cadaba farinosa, Calliandra spp, Camellia sinensis, Canna indica, Capsicum spp., Cassia spp., Centroema pubescens, Chacoomeles spp., Cinnamomum cassia, Coffea arabica, Colophospermum mopane, Coronillia varia, Cotoneaster serotina, Crataegus spp., Cucumis spp., Cupressus spp., Cyathea dealbata, Cydonia oblonga, Cryptomeria japonica, Cymbopogon spp., Cynthea dealbata, Cydonia oblonga, Dalbergia monetaria, Davallia divaricata, Desmodium spp., Dicksonia squarosa, Dibeteropogon amplectens, Dioclea spp, Dolichos spp., Dorycnium rectum, Echinochloa pyramidalis, Ehraffia spp., Eleusine coracana, Eragrestis spp., Erythrina spp., Eucalypfus spp., Euclea schimperi, Eulalia vi/losa, Pagopyrum spp., Feijoa sellowlana, Fragaria spp., Flemingia spp, Freycinetia banksli, Geranium thunbergii, GinAgo biloba, Glycine javanica, Gliricidia spp, Gossypium hirsutum, Grevillea spp., Guibourtia coleosperma, Hedysarum spp., Hemaffhia altissima, Heteropogon contoffus, Hordeum vulgare, Hyparrhenia rufa, Hypericum erectum, Hypeffhelia dissolute, Indigo incamata, Iris spp., Leptarrhena pyrolifolia, Lespediza spp., Lettuca spp., Leucaena leucocephala, Loudetia simplex, Lotonus bainesli, Lotus spp., Macrotyloma axillare, Malus spp., Manihot esculenta, Medicago saliva, Metasequoia glyptostroboides, Musa sapientum, Nicotianum spp., Onobrychis spp., Ornithopus spp., Oryza spp., Peltophorum africanum, Pennisetum spp., Persea gratissima, Petunia spp., Phaseolus spp., Phoenix canariensis, Phormium cookianum, Photinia spp., Picea glauca, Pinus spp., Pisum sativam, Podocarpus totara, Pogonarthria fleckii, Pogonaffhria squarrosa, Populus spp., Prosopis cineraria, Pseudotsuga menziesii, Pterolobium stellatum, Pyrus communis, Quercus spp., Rhaphiolepsis umbellata, Rhopalostylis sapida, Rhus natalensis, Ribes grossularia, Ribes spp., Robinia pseudoacacia, Rosa spp., Rubus spp., Salix spp., Schyzachyrium sanguineum, Sciadopitys vefficillata, Sequoia sempervirens, Sequoiadendron giganteum, Sorghum bicolor, Spinacia spp., Sporobolus fimbriatus, Stiburus alopecuroides, Stylosanthos humilis, Tadehagi spp, Taxodium distichum, Themeda triandra, Trifolium spp., Triticum spp., Tsuga heterophylla, Vaccinium spp., Vicia spp., Vitis vinifera, Watsonia pyramidata, Zantedeschia aethiopica, Zea mays, amaranth, artichoke, asparagus, broccoli, Brussels sprouts, cabbage, canola, carrot, cauliflower, celery, collard greens, flax, kale, lentil, oilseed rape, okra, onion, potato, rice, soybean, straw, sugar beet, sugar cane, sunflower, tomato, squash tea, maize, wheat, barley, rye, oat, peanut, pea, lentil and alfalfa, cotton, rapeseed, canola, pepper, sunflower, tobacco, eggplant, eucalyptus, a tree, an ornamental plant, a perennial grass and a forage crop. Alternatively algae and other non-Viridiplantae can be used for the methods of the present invention.
[0119] According to some embodiments of the invention, the plant used by the method of the invention is a crop plant such as rice, maize, wheat, barley, peanut, potato, sesame, olive tree, palm oil, banana, soybean, sunflower, canola, sugarcane, alfalfa, millet, leguminosae (bean, pea), flax, lupinus, rapeseed, tobacco, poplar and cotton.
[0120] According to some embodiments of the invention, the plant is a tomato or a banana.
[0121] According to some embodiments of the invention, expressing the exogenous polynucleotide of the invention within the plant is effected by introducing into a cell of the plant (e.g., transforming one or more cells of the plant) an exogenous polynucleotide encoding the HXK under a cis-acting regulatory element for driving expression of the HXK in a guard-cell specific manner, followed by generating a mature plant from the transformed cells and cultivating the mature plant under conditions suitable for expressing the exogenous polynucleotide within the mature plant.
[0122] Thus, there is provided a plant expression construct comprising a nucleic acid sequence encoding a hexokinase under a transcriptional control of a guard cell-specific cis-acting regulatory element and methods which make use of same.
[0123] There is also provided a method of decreasing plant stomata conductance, the method comprising introducing into a cell of a plant the above-described nucleic acid construct, thereby decreasing the stomata conductance of the plant.
[0124] Alternatively or additionally there is provided a method of increasing water use efficiency of a plant, the method comprising introducing into a cell of the plant the above-described nucleic acid construct, thereby increasing water use efficiency of the plant.
[0125] Alternatively or additionally there is provided a method of increasing tolerance of a plant to drought, salinity or temperature stress, the method comprising introducing into a cell of the plant the above-described nucleic acid construct, thereby increasing tolerance of the plant to drought, salinity or temperature stress.
[0126] Alternatively or additionally there is provided a method of increasing biotic stress tolerance of a plant, the method comprising introducing into a cell of the plant the above-described nucleic acid construct, thereby increasing biotic stress tolerance of the plant.
[0127] Alternatively or additionally there is provided a method of increasing biomass, vigor or yield of a plant, the method comprising introducing into a cell of the plant the nucleic acid construct, thereby increasing the biomass, vigor or yield of the plant
[0128] According to some embodiments of the invention, the transformation is effected by introducing to the plant cell a nucleic acid construct which includes the exogenous polynucleotide of some embodiments of the invention encoding the HXK (as described above) and a guard cell-specific cis-acting regulatory element. Further details of suitable transformation approaches are provided hereinbelow.
[0129] As used herein "guard-cell specific cis-acting regulatory element" refers to the ability of a transcriptional element to drive expression of the nucleic acid sequence under its regulation (e.g., HXK) only in guard cells, leaving the rest of the tissues in the plant unmodified by transgene expression (e.g., more than 90% of the mRNA is expressed in the tissue of interest, as detected by RT-PCR). Tissue-specific cis-acting regulatory elements may be induced by endogenous or exogenous factors, so they can be classified as inducible promoters as well. In other cases they are constitutively expressed.
[0130] A coding nucleic acid sequence (e.g., HXK) is "operably linked" to a regulatory sequence (e.g., guard-cell specific promoter) if the regulatory sequence is capable of exerting a regulatory effect on the coding sequence linked thereto.
[0131] According to some embodiments of the invention the cis-acting regulatory element is a promoter.
[0132] As used herein, the term "promoter" refers to a region of DNA which lies upstream of the transcriptional initiation site of a gene to which RNA polymerase binds to initiate transcription of RNA. The promoter controls where (e.g., which portion of a plant) and/or when (e.g., at which stage or condition in the lifetime of an organism) the gene is expressed.
[0133] Examples of guard-cell specific promoters include, but are not limited to the promoters listed in Table 2 below and the KST1 promoter used in the Examples section (SEQ ID NO: 108).
TABLE-US-00002 TABLE 2 Verification Promoter Species Accession n. method Ref. Comments 1 AtMYB61 Arabidopsis AT1G09540 GFP (Liang et al., 2005) Specific promoter thaliana (SEQ ID NO: 95) expression in GC 2 At1g22690- Arabidopsis At1g22690 GFP based (Yang et al., 2008) Specific promoter thaliana (SEQ ID NO: 96) calcium expression in GC (pGC1) FRET reporter/GUS 3 AtMYB60 Arabidopsis At1g08810 GUS, GFP (Cominelli et al., 2005; Specific promoter thaliana (SEQ ID NO: 97) Galbiati et al., 2008; expression in GC Cominelli et al., 2011) 4 R2R3 MYB60 Vitis ACF21938 GUS (Galbiati et al., 2011) Specific transcription vinifera (SEQ ID NO: 98) expression in GC factor promoter L. 5 HIC (High Arabidopsis AT2G46720 GUS (Gray et al., 2000) Specific carbon dioxide) thaliana (SEQ ID NO: 99) expression in GC promoter 6 CYTOCHROME Arabidopsis At4g00360 GFP (Francia et al.,2008; Specific P450 86A2 thaliana (SEQ ID NO: 100) Galbiati et al., 2008) expression in GC (CYP86A2) mono- oxygenase promoter (pCYP) 7 ADP-glucose Solanum X75017 GUS (Muller-Rober et al., 0.3 Kb 5'proximal pyrophosphory- tuberosum (Promoter seq.) 1994) promoter - lase (AGPase) (SEQ ID NO: 101) exclusive GC Promoter expression 8 KAT1 promoter Arabidopsis AT5G46240 GUS (Nakamura et al., 1995) Specific thaliana (gene), U25088 expression in GC. (promoter + However, was gene seq.) detected also in (SEQ ID NO: 102) vascular tissue of roots 9 Myrosinase- Arabidopsis At5g26000 GUS, GFP (Husebye et al., 2002) Specific Thioglucoside thaliana (SEQ ID NO: 103) expression in GC. glucohydrolase Distinct 1 (TGG1) expression in promoter phloem 10 rha1 promoter Arabidopsis AT5G45130 GUS (Terryn et al., 1993) Mainly expressed thaliana (SEQ ID NO: 104) (non-specific) in GC 11 AtCHX20 Arabidopsis AT3G53720 GUS (Padmanaban et al., Specific promoter thaliana (SEQ ID NO: 105) 2007) expression in GC GC--guard cell. GFP--green fluorescence protein. GUS--β-glucoronidase reporter gene.
[0134] The nucleic acid construct of some embodiments of the invention can further include an appropriate selectable marker and/or an origin of replication. According to some embodiments of the invention, the nucleic acid construct utilized is a shuttle vector, which can propagate both in E. coli (wherein the construct comprises an appropriate selectable marker and origin of replication) and be compatible with propagation in cells. The construct according to the present invention can be, for example, a plasmid, a bacmid, a phagemid, a cosmid, a phage, a virus or an artificial chromosome.
[0135] The nucleic acid construct of some embodiments of the invention can be utilized to stably or transiently transform plant cells. In stable transformation, the exogenous polynucleotide is integrated into the plant genome and as such it represents a stable and inherited trait. In transient transformation, the exogenous polynucleotide is expressed by the cell transformed but it is not integrated into the genome and as such it represents a transient trait.
[0136] There are various methods of introducing foreign genes into both monocotyledonous and dicotyledonous plants (Potrykus, I., Annu. Rev. Plant. Physiol., Plant. Mol. Biol. (1991) 42:205-225; Shimamoto et al., Nature (1989) 338:274-276).
[0137] The principle methods of causing stable integration of exogenous DNA into plant genomic DNA include two main approaches:
[0138] (i) Agrobacterium-mediated gene transfer: Klee et al. (1987) Annu. Rev. Plant Physiol. 38:467-486; Klee and Rogers in Cell Culture and Somatic Cell Genetics of Plants, Vol. 6, Molecular Biology of Plant Nuclear Genes, eds. Schell, J., and Vasil, L. K., Academic Publishers, San Diego, Calif. (1989) p. 2-25; Gatenby, in Plant Biotechnology, eds. Kung, S, and Arntzen, C. J., Butterworth Publishers, Boston, Mass. (1989) p. 93-112.
[0139] (ii) Direct DNA uptake: Paszkowski et al., in Cell Culture and Somatic Cell Genetics of Plants, Vol. 6, Molecular Biology of Plant Nuclear Genes eds. Schell, J., and Vasil, L. K., Academic Publishers, San Diego, Calif. (1989) p. 52-68; including methods for direct uptake of DNA into protoplasts, Toriyama, K. et al. (1988) Bio/Technology 6:1072-1074. DNA uptake induced by brief electric shock of plant cells: Zhang et al. Plant Cell Rep. (1988) 7:379-384. Fromm et al. Nature (1986) 319:791-793. DNA injection into plant cells or tissues by particle bombardment, Klein et al. Bio/Technology (1988) 6:559-563; McCabe et al. Bio/Technology (1988) 6:923-926; Sanford, Physiol. Plant. (1990) 79:206-209; by the use of micropipette systems: Neuhaus et al., Theor. Appl. Genet. (1987) 75:30-36; Neuhaus and Spangenberg, Physiol. Plant. (1990) 79:213-217; glass fibers or silicon carbide whisker transformation of cell cultures, embryos or callus tissue, U.S. Pat. No. 5,464,765 or by the direct incubation of DNA with germinating pollen, DeWet et al. in Experimental Manipulation of Ovule Tissue, eds. Chapman, G. P. and Mantell, S. H. and Daniels, W. Longman, London, (1985) p. 197-209; and Ohta, Proc. Natl. Acad. Sci. USA (1986) 83:715-719.
[0140] The Agrobacterium system includes the use of plasmid vectors that contain defined DNA segments that integrate into the plant genomic DNA. Methods of inoculation of the plant tissue vary depending upon the plant species and the Agrobacterium delivery system. A widely used approach is the leaf disc procedure which can be performed with any tissue explant that provides a good source for initiation of whole plant differentiation. See, e.g., Horsch et al. in Plant Molecular Biology Manual A5, Kluwer Academic Publishers, Dordrecht (1988) p. 1-9. A supplementary approach employs the Agrobacterium delivery system in combination with vacuum infiltration. The Agrobacterium system is especially viable in the creation of transgenic dicotyledonous plants.
[0141] There are various methods of direct DNA transfer into plant cells. In electroporation, the protoplasts are briefly exposed to a strong electric field. In microinjection, the DNA is mechanically injected directly into the cells using very small micropipettes. In microparticle bombardment, the DNA is adsorbed on microprojectiles such as magnesium sulfate crystals or tungsten particles, and the microprojectiles are physically accelerated into cells or plant tissues.
[0142] Following stable transformation plant propagation is exercised. The most common method of plant propagation is by seed. Regeneration by seed propagation, however, has the deficiency that due to heterozygosity there is a lack of uniformity in the crop, since seeds are produced by plants according to the genetic variances governed by Mendelian rules. Basically, each seed is genetically different and each will grow with its own specific traits. Therefore, it is preferred that the transformed plant be produced such that the regenerated plant has the identical traits and characteristics of the parent transgenic plant. Therefore, it is preferred that the transformed plant be regenerated by micropropagation which provides a rapid, consistent reproduction of the transformed plants.
[0143] Micropropagation is a process of growing new generation plants from a single piece of tissue that has been excised from a selected parent plant or cultivar. This process permits the mass reproduction of plants having the preferred tissue expressing the fusion protein. The new generation plants which are produced are genetically identical to, and have all of the characteristics of, the original plant. Micropropagation allows mass production of quality plant material in a short period of time and offers a rapid multiplication of selected cultivars in the preservation of the characteristics of the original transgenic or transformed plant. The advantages of cloning plants are the speed of plant multiplication and the quality and uniformity of plants produced.
[0144] Micropropagation is a multi-stage procedure that requires alteration of culture medium or growth conditions between stages. Thus, the micropropagation process involves four basic stages: Stage one, initial tissue culturing; stage two, tissue culture multiplication; stage three, differentiation and plant formation; and stage four, greenhouse culturing and hardening. During stage one, initial tissue culturing, the tissue culture is established and certified contaminant-free. During stage two, the initial tissue culture is multiplied until a sufficient number of tissue samples are produced to meet production goals. During stage three, the tissue samples grown in stage two are divided and grown into individual plantlets. At stage four, the transformed plantlets are transferred to a greenhouse for hardening where the plants' tolerance to light is gradually increased so that it can be grown in the natural environment.
[0145] According to some embodiments of the invention, the transgenic plants are generated by transient transformation of leaf cells, meristematic cells or the whole plant.
[0146] Transient transformation can be effected by any of the direct DNA transfer methods described above or by viral infection using modified plant viruses.
[0147] Viruses that have been shown to be useful for the transformation of plant hosts include CaMV, Tobacco mosaic virus (TMV), brome mosaic virus (BMV) and Bean Common Mosaic Virus (BV or BCMV). Transformation of plants using plant viruses is described in U.S. Pat. No. 4,855,237 (bean golden mosaic virus; BGV), EP-A 67,553 (TMV), Japanese Published Application No. 63-14693 (TMV), EPA 194,809 (BV), EPA 278,667 (BV); and Gluzman, Y. et al., Communications in Molecular Biology: Viral Vectors, Cold Spring Harbor Laboratory, New York, pp. 172-189 (1988). Pseudovirus particles for use in expressing foreign DNA in many hosts, including plants are described in WO 87/06261.
[0148] According to some embodiments of the invention, the virus used for transient transformations is avirulent and thus is incapable of causing severe symptoms such as reduced growth rate, mosaic, ring spots, leaf roll, yellowing, streaking, pox formation, tumor formation and pitting. A suitable avirulent virus may be a naturally occurring avirulent virus or an artificially attenuated virus. Virus attenuation may be effected by using methods well known in the art including, but not limited to, sub-lethal heating, chemical treatment or by directed mutagenesis techniques such as described, for example, by Kurihara and Watanabe (Molecular Plant Pathology 4:259-269, 2003), Gal-on et al. (1992), Atreya et al. (1992) and Huet et al. (1994).
[0149] Suitable virus strains can be obtained from available sources such as, for example, the American Type culture Collection (ATCC) or by isolation from infected plants. Isolation of viruses from infected plant tissues can be effected by techniques well known in the art such as described, for example by Foster and Tatlor, Eds. "Plant Virology Protocols: From Virus Isolation to Transgenic Resistance (Methods in Molecular Biology (Humana Pr), Vol 81)", Humana Press, 1998. Briefly, tissues of an infected plant believed to contain a high concentration of a suitable virus, preferably young leaves and flower petals, are ground in a buffer solution (e.g., phosphate buffer solution) to produce a virus infected sap which can be used in subsequent inoculations.
[0150] Construction of plant RNA viruses for the introduction and expression of non-viral exogenous polynucleotide sequences in plants is demonstrated by the above references as well as by Dawson, W. O. et al., Virology (1989) 172:285-292; Takamatsu et al. EMBO J. (1987) 6:307-311; French et al. Science (1986) 231:1294-1297; Takamatsu et al. FEBS Letters (1990) 269:73-76; and U.S. Pat. No. 5,316,931.
[0151] When the virus is a DNA virus, suitable modifications can be made to the virus itself. Alternatively, the virus can first be cloned into a bacterial plasmid for ease of constructing the desired viral vector with the foreign DNA. The virus can then be excised from the plasmid. If the virus is a DNA virus, a bacterial origin of replication can be attached to the viral DNA, which is then replicated by the bacteria. Transcription and translation of this DNA will produce the coat protein which will encapsidate the viral DNA. If the virus is an RNA virus, the virus is generally cloned as a cDNA and inserted into a plasmid. The plasmid is then used to make all of the constructions. The RNA virus is then produced by transcribing the viral sequence of the plasmid and translation of the viral genes to produce the coat protein(s) which encapsidate the viral RNA.
[0152] In one embodiment, a plant viral polynucleotide is provided in which the native coat protein coding sequence has been deleted from a viral polynucleotide, a non-native plant viral coat protein coding sequence and a non-native promoter, preferably the subgenomic promoter of the non-native coat protein coding sequence, capable of expression in the plant host, packaging of the recombinant plant viral polynucleotide, and ensuring a systemic infection of the host by the recombinant plant viral polynucleotide, has been inserted. Alternatively, the coat protein gene may be inactivated by insertion of the non-native polynucleotide sequence within it, such that a protein is produced. The recombinant plant viral polynucleotide may contain one or more additional non-native subgenomic promoters. Each non-native subgenomic promoter is capable of transcribing or expressing adjacent genes or polynucleotide sequences in the plant host and incapable of recombination with each other and with native subgenomic promoters. Non-native (foreign) polynucleotide sequences may be inserted adjacent the native plant viral subgenomic promoter or the native and a non-native plant viral subgenomic promoters if more than one polynucleotide sequence is included. The non-native polynucleotide sequences are transcribed or expressed in the host plant under control of the subgenomic promoter to produce the desired products.
[0153] In a second embodiment, a recombinant plant viral polynucleotide is provided as in the first embodiment except that the native coat protein coding sequence is placed adjacent one of the non-native coat protein subgenomic promoters instead of a non-native coat protein coding sequence.
[0154] In a third embodiment, a recombinant plant viral polynucleotide is provided in which the native coat protein gene is adjacent its subgenomic promoter and one or more non-native subgenomic promoters have been inserted into the viral polynucleotide. The inserted non-native subgenomic promoters are capable of transcribing or expressing adjacent genes in a plant host and are incapable of recombination with each other and with native subgenomic promoters. Non-native polynucleotide sequences may be inserted adjacent the non-native subgenomic plant viral promoters such that the sequences are transcribed or expressed in the host plant under control of the subgenomic promoters to produce the desired product.
[0155] In a fourth embodiment, a recombinant plant viral polynucleotide is provided as in the third embodiment except that the native coat protein coding sequence is replaced by a non-native coat protein coding sequence.
[0156] The viral vectors are encapsidated by the coat proteins encoded by the recombinant plant viral polynucleotide to produce a recombinant plant virus. The recombinant plant viral polynucleotide or recombinant plant virus is used to infect appropriate host plants. The recombinant plant viral polynucleotide is capable of replication in the host, systemic spread in the host, and transcription or expression of foreign gene(s) (exogenous polynucleotide) in the host to produce the desired protein.
[0157] Techniques for inoculation of viruses to plants may be found in Foster and Taylor, eds. "Plant Virology Protocols: From Virus Isolation to Transgenic Resistance (Methods in Molecular Biology (Humana Pr), Vol 81)", Humana Press, 1998; Maramorosh and Koprowski, eds. "Methods in Virology" 7 vols, Academic Press, New York 1967-1984; Hill, S. A. "Methods in Plant Virology", Blackwell, Oxford, 1984; Walkey, D. G. A. "Applied Plant Virology", Wiley, New York, 1985; and Kado and Agrawa, eds. "Principles and Techniques in Plant Virology", Van Nostrand-Reinhold, New York.
[0158] In addition to the above, the polynucleotide of the present invention can also be introduced into a chloroplast genome thereby enabling chloroplast expression.
[0159] A technique for introducing exogenous polynucleotide sequences to the genome of the chloroplasts is known. This technique involves the following procedures. First, plant cells are chemically treated so as to reduce the number of chloroplasts per cell to about one. Then, the exogenous polynucleotide is introduced via particle bombardment into the cells with the aim of introducing at least one exogenous polynucleotide molecule into the chloroplasts. The exogenous polynucleotides selected such that it is integratable into the chloroplast's genome via homologous recombination which is readily effected by enzymes inherent to the chloroplast. To this end, the exogenous polynucleotide includes, in addition to a gene of interest, at least one polynucleotide stretch which is derived from the chloroplast's genome. In addition, the exogenous polynucleotide includes a selectable marker, which serves by sequential selection procedures to ascertain that all or substantially all of the copies of the chloroplast genomes following such selection will include the exogenous polynucleotide. Further details relating to this technique are found in U.S. Pat. No. 4,945,050; and U.S. Pat. No. 5,693,507 which are incorporated herein by reference. A polypeptide can thus be produced by the protein expression system of the chloroplast and become integrated into the chloroplast's inner membrane.
[0160] According to some embodiments of the invention, the method further comprising growing the plant expressing the exogenous polynucleotide under the biotic or abiotic stress (e.g., drought, water deprivation or temperature stress). Thus, the invention encompasses (transgenic) plants, parts thereof or plant cells, exogenously expressing the polynucleotide(s) or the nucleic acid constructs of the invention.
[0161] Once expressed within the plant cell or the entire plant, the level of the polypeptide encoded by the exogenous polynucleotide can be determined by methods well known in the art such as, activity assays, Western blots using antibodies capable of specifically binding the polypeptide, Enzyme-Linked Immuno Sorbent Assay (ELISA), radio-immuno-assays (RIA), immunohistochemistry, immunocytochemistry, immunofluorescence and the like.
[0162] Methods of determining the level in the plant of the RNA transcribed from the exogenous polynucleotide are well known in the art and include, for example, Northern blot analysis, reverse transcription polymerase chain reaction (RT-PCR) analysis (including quantitative, semi-quantitative or real-time RT-PCR) and RNA-in situ hybridization.
[0163] The effect of the expressed HXK on plant stomata conductance (e.g., manifested by aperture), water use efficiency, water use efficiency and/or photosynthesis can be qualified using methods which are well known in the art. Stomata functionality assays are described in length in the Examples section which follows.
[0164] The effect of the exogenous polynucleotide encoding the HXK on abiotic stress tolerance can be determined using known methods such as detailed below and in the Examples section which follows.
[0165] Abiotic stress tolerance--Transformed (i.e., expressing the HXK) and non-transformed (wild type) plants are exposed to biotic or an abiotic stress condition, such as water deprivation or suboptimal temperature (low temperature, high temperature).
[0166] Cold stress tolerance--To analyze cold stress, mature (25 day old) plants are transferred to 4° C. chambers for 1 or 2 weeks, with constitutive light. Later on plants are moved back to greenhouse. Two weeks later damages from chilling period, resulting in growth retardation and other phenotypes, are compared between both control and transgenic plants, by measuring plant weight (wet and dry), and by comparing growth rates measured as time to flowering, plant size, yield, and the like.
[0167] Heat stress tolerance--Heat stress tolerance is achieved by exposing the plants to temperatures above 34° C. for a certain period. Plant tolerance is examined after transferring the plants back to 22° C. for recovery and evaluation after 5 days relative to internal controls (non-transgenic plants) or plants not exposed to neither cold or heat stress.
[0168] Water use efficiency--can be determined as the biomass produced per unit transpiration. To analyze WUE, leaf relative water content can be measured in control and transgenic plants. Fresh weight (FW) is immediately recorded; then leaves are soaked for 8 hours in distilled water at room temperature in the dark, and the turgid weight (TW) is recorded. Total dry weight (DW) is recorded after drying the leaves at 60° C. to a constant weight. Relative water content (RWC) is calculated.
[0169] Salinity tolerance assay--Transgenic plants with tolerance to high salt concentrations are expected to exhibit better germination, seedling vigor or growth in high salt. Salt stress can be effected in many ways such as, for example, by irrigating the plants with a hyperosmotic solution, by cultivating the plants hydroponically in a hyperosmotic growth solution (e.g., Hoagland solution), or by culturing the plants in a hyperosmotic growth medium [e.g., 50% Murashige-Skoog medium (MS medium)]. Since different plants vary considerably in their tolerance to salinity, the salt concentration in the irrigation water, growth solution, or growth medium can be adjusted according to the specific characteristics of the specific plant cultivar or variety, so as to inflict a mild or moderate effect on the physiology and/or morphology of the plants (for guidelines as to appropriate concentration see, Bernstein and Kafkafi, Root Growth Under Salinity Stress In: Plant Roots, The Hidden Half 3rd ed. Waisel Y, Eshel A and Kafkafi U. (editors) Marcel Dekker Inc., New York, 2002, and reference therein).
[0170] For example, a salinity tolerance test can be performed by irrigating plants at different developmental stages with increasing concentrations of sodium chloride (for example 50 mM, 100 mM, 200 mM, 400 mM NaCl) applied from the bottom and from above to ensure even dispersal of salt. Following exposure to the stress condition the plants are frequently monitored until substantial physiological and/or morphological effects appear in wild type plants. Thus, the external phenotypic appearance, degree of wilting and overall success to reach maturity and yield progeny are compared between control and transgenic plants.
[0171] Quantitative parameters of tolerance measured include, but are not limited to, the average wet and dry weight, growth rate, leaf size, leaf coverage (overall leaf area), the weight of the seeds yielded, the average seed size and the number of seeds produced per plant. Transformed plants not exhibiting substantial physiological and/or morphological effects, or exhibiting higher biomass than wild-type plants, are identified as abiotic stress tolerant plants.
[0172] Osmotic tolerance test--Osmotic stress assays (including sodium chloride and mannitol assays) are conducted to determine if an osmotic stress phenotype was sodium chloride-specific or if it was a general osmotic stress related phenotype. Plants which are tolerant to osmotic stress may have more tolerance to drought and/or freezing. For salt and osmotic stress germination experiments, the medium is supplemented for example with 50 mM, 100 mM, 200 mM NaCl or 100 mM, 200 mM NaCl, 400 mM mannitol.
[0173] The effect of the transgene on plant's vigor, growth rate, biomass, yield and/or oil content can be determined using known methods.
[0174] Plant vigor--The plant vigor can be calculated by the increase in growth parameters such as leaf area, fiber length, rosette diameter, plant fresh weight and the like per time.
[0175] Growth rate--The growth rate can be measured using digital analysis of growing plants. For example, images of plants growing in greenhouse on plot basis can be captured every 3 days and the rosette area can be calculated by digital analysis. Rosette area growth is calculated using the difference of rosette area between days of sampling divided by the difference in days between samples.
[0176] As mentioned, the present teachings are also directed at downregulating HXK activity or expression in a guard cell specific manner. This is effected to increase plant dehydration where needed. For example when there is a need to accelerate defoliation prior or after harvesting such as in cotton and other crops, or for dehydration of leaves and stems for straw for instance.
[0177] Downregulation (gene silencing) of the transcription or translation product of an endogenous HXK in a guard-cell specific manner can be achieved by co-suppression, antisense suppression, RNA interference and ribozyme molecules under the above mentioned cis-acting regulatory element active specifically in a guard cell.
[0178] Thus, there is provided a plant expression construct comprising a nucleic acid sequence encoding a nucleic acid agent for silencing expression of a hexokinase, wherein expression of said nucleic acid agent is under a transcriptional control of a guard cell-specific cis-acting regulatory element (as described above).
[0179] Co-suppression (sense suppression)--Inhibition of the endogenous gene can be achieved by co-suppression, using an RNA molecule (or an expression vector encoding same) which is in the sense orientation with respect to the transcription direction of the endogenous gene. The polynucleotide used for co-suppression may correspond to all or part of the sequence encoding the endogenous polypeptide and/or to all or part of the 5' and/or 3' untranslated region of the endogenous transcript; it may also be an unpolyadenylated RNA; an RNA which lacks a 5' cap structure; or an RNA which contains an unsplicable intron. In some embodiments, the polynucleotide used for co-suppression is designed to eliminate the start codon of the endogenous polynucleotide so that no protein product will be translated. Methods of co-suppression using a full-length cDNA sequence as well as a partial cDNA sequence are known in the art (see, for example, U.S. Pat. No. 5,231,020).
[0180] According to some embodiments of the invention, downregulation of the endogenous gene is performed using an amplicon expression vector which comprises a plant virus-derived sequence that contains all or part of the target gene but generally not all of the genes of the native virus. The viral sequences present in the transcription product of the expression vector allow the transcription product to direct its own replication. The transcripts produced by the amplicon may be either sense or antisense relative to the target sequence [see for example, Angell and Baulcombe, (1997) EMBO J. 16:3675-3684; Angell and Baulcombe, (1999) Plant J. 20:357-362, and U.S. Pat. No. 6,646,805, each of which is herein incorporated by reference].
[0181] Antisense suppression--Antisense suppression can be performed using an antisense polynucleotide or an expression vector which is designed to express an RNA molecule complementary to all or part of the messenger RNA (mRNA) encoding the endogenous polypeptide and/or to all or part of the 5' and/or 3' untranslated region of the endogenous gene. Over expression of the antisense RNA molecule can result in reduced expression of the native (endogenous) gene. The antisense polynucleotide may be fully complementary to the target sequence (i.e., 100% identical to the complement of the target sequence) or partially complementary to the target sequence (i.e., less than 100% identical, e.g., less than 90%, less than 80% identical to the complement of the target sequence). Antisense suppression may be used to inhibit the expression of multiple proteins in the same plant (see e.g., U.S. Pat. No. 5,942,657). In addition, portions of the antisense nucleotides may be used to disrupt the expression of the target gene. Generally, sequences of at least about 50 nucleotides, at least about 100 nucleotides, at least about 200 nucleotides, at least about 300, at least about 400, at least about 450, at least about 500, at least about 550, or greater may be used. Methods of using antisense suppression to inhibit the expression of endogenous genes in plants are described, for example, in Liu, et al., (2002) Plant Physiol. 129:1732-1743 and U.S. Pat. Nos. 5,759,829 and 5,942,657, each of which is herein incorporated by reference. Efficiency of antisense suppression may be increased by including a poly-dT region in the expression cassette at a position 3' to the antisense sequence and 5' of the polyadenylation signal [See, U.S. Patent Publication No. 20020048814, herein incorporated by reference].
[0182] RNA interference--RNA interference can be achieved using a polynucleotide, which can anneal to itself and form a double stranded RNA having a stem-loop structure (also called hairpin structure), or using two polynucleotides, which form a double stranded RNA.
[0183] For hairpin RNA (hpRNA) interference, the expression vector is designed to express an RNA molecule that hybridizes to itself to form a hairpin structure that comprises a single-stranded loop region and a base-paired stem.
[0184] In some embodiments of the invention, the base-paired stem region of the hpRNA molecule determines the specificity of the RNA interference. In this configuration, the sense sequence of the base-paired stem region may correspond to all or part of the endogenous mRNA to be downregulated, or to a portion of a promoter sequence controlling expression of the endogenous gene to be inhibited; and the antisense sequence of the base-paired stem region is fully or partially complementary to the sense sequence. Such hpRNA molecules are highly efficient at inhibiting the expression of endogenous genes, in a manner which is inherited by subsequent generations of plants [See, e.g., Chuang and Meyerowitz, (2000) Proc. Natl. Acad. Sci. USA 97:4985-4990; Stoutjesdijk, et al., (2002) Plant Physiol. 129:1723-1731; and Waterhouse and Helliwell, (2003) Nat. Rev. Genet. 4:29-38; Chuang and Meyerowitz, (2000) Proc. Natl. Acad. Sci. USA 97:4985-4990; Pandolfini et al., BMC Biotechnology 3:7; Panstruga, et al., (2003) Mol. Biol. Rep. 30:135-140; and U.S. Patent Publication No. 2003/0175965; each of which is incorporated by reference].
[0185] According to some embodiments of the invention, the sense sequence of the base-paired stem is from about 10 nucleotides to about 2,500 nucleotides in length, e.g., from about 10 nucleotides to about 500 nucleotides, e.g., from about 15 nucleotides to about 300 nucleotides, e.g., from about 20 nucleotides to about 100 nucleotides, e.g., or from about 25 nucleotides to about 100 nucleotides.
[0186] According to some embodiments of the invention, the antisense sequence of the base-paired stem may have a length that is shorter, the same as, or longer than the length of the corresponding sense sequence.
[0187] According to some embodiments of the invention, the loop portion of the hpRNA can be from about 10 nucleotides to about 500 nucleotides in length, for example from about 15 nucleotides to about 100 nucleotides, from about 20 nucleotides to about 300 nucleotides or from about 25 nucleotides to about 400 nucleotides in length.
[0188] According to some embodiments of the invention, the loop portion of the hpRNA can include an intron (ihpRNA), which is capable of being spliced in the host cell. The use of an intron minimizes the size of the loop in the hairpin RNA molecule following splicing and thus increases efficiency of the interference [See, for example, Smith, et al., (2000) Nature 407:319-320; Wesley, et al., (2001) Plant J. 27:581-590; Wang and Waterhouse, (2001) Curr. Opin. Plant Biol. 5:146-150; Helliwell and Waterhouse, (2003) Methods 30:289-295; Brummell, et al. (2003) Plant J. 33:793-800; and U.S. Patent Publication No. 2003/0180945; WO 98/53083; WO 99/32619; WO 98/36083; WO 99/53050; US 20040214330; US 20030180945; U.S. Pat. No. 5,034,323; U.S. Pat. No. 6,452,067; U.S. Pat. No. 6,777,588; U.S. Pat. No. 6,573,099 and U.S. Pat. No. 6,326,527; each of which is herein incorporated by reference].
[0189] In some embodiments of the invention, the loop region of the hairpin RNA determines the specificity of the RNA interference to its target endogenous RNA. In this configuration, the loop sequence corresponds to all or part of the endogenous messenger RNA of the target gene. See, for example, WO 02/00904; Mette, et al., (2000) EMBO J. 19:5194-5201; Matzke, et al., (2001) Curr. Opin. Genet. Devel. 11:221-227; Scheid, et al., (2002) Proc. Natl. Acad. Sci., USA 99:13659-13662; Aufsaftz, et al., (2002) Proc. Nat'l. Acad. Sci. 99(4):16499-16506; Sijen, et al., Curr. Biol. (2001) 11:436-440), each of which is incorporated herein by reference.
[0190] For double-stranded RNA (dsRNA) interference, the sense and antisense RNA molecules can be expressed in the same cell from a single expression vector (which comprises sequences of both strands) or from two expression vectors (each comprising the sequence of one of the strands). Methods for using dsRNA interference to inhibit the expression of endogenous plant genes are described in Waterhouse, et al., (1998) Proc. Natl. Acad. Sci. USA 95:13959-13964; and WO 99/49029, WO 99/53050, WO 99/61631, and WO 00/49035; each of which is herein incorporated by reference.
[0191] According to some embodiments of the invention, RNA intereference is effected using an expression vector designed to express an RNA molecule that is modeled on an endogenous micro RNAs (miRNA) gene. Micro RNAs (miRNAs) are regulatory agents consisting of about 22 ribonucleotides and highly efficient at inhibiting the expression of endogenous genes [Javier, et al., (2003) Nature 425:257-263]. The miRNA gene encodes an RNA that forms a hairpin structure containing a 22-nucleotide sequence that is complementary to the endogenous target gene.
[0192] Thus, the present teachings provide for a transgenic plant or a part thereof comprising the plant expression construct as described herein as well as isolated plant cell or a plant cell culture comprising the plant expression construct as described herein.
[0193] The present teachings also relate to processed products produced from the plants, plant parts or plant cells of the present invention. Such processed products relate to food, animal feed, beverages, construction material, biofuel, biodiesel, oils, sauces, pastes, pastries, meal and the like.
[0194] It is expected that during the life of a patent maturing from this application many relevant hexokinases and guard cell specific cis-acting regulatory elements will be developed and the scope of the terms used herein are intended to include all such new technologies a priori.
[0195] As used herein the term "about" refers to ±10%.
[0196] The terms "comprises", "comprising", "includes", "including", "having" and their conjugates mean "including but not limited to".
[0197] The term "consisting of" means "including and limited to".
[0198] The term "consisting essentially of" means that the composition, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients, steps and/or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.
[0199] As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof.
[0200] Throughout this application, various embodiments of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.
[0201] Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases "ranging/ranges between" a first indicate number and a second indicate number and "ranging/ranges from" a first indicate number "to" a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.
[0202] As used herein the term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.
[0203] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.
[0204] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.
EXAMPLES
[0205] Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non limiting fashion.
[0206] Generally, the nomenclature used herein and the laboratory procedures utilized in the present invention include molecular, biochemical, cellular and recombinant DNA techniques. Such techniques are thoroughly explained in the literature. See, for example, "Molecular Cloning: A laboratory Manual" Sambrook et al., (1989); "Current Protocols in Molecular Biology" Volumes I-III Ausubel, R. M., ed. (1994); Ausubel et al., "Current Protocols in Molecular Biology", John Wiley and Sons, Baltimore, Md. (1989); Perbal, "A Practical Guide to Molecular Cloning", John Wiley & Sons, New York (1988); Watson et al., "Recombinant DNA", Scientific American Books, New York; Birren et al. (eds); methodologies as set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and 5,272,057; "Cell Biology: A Laboratory Handbook", Volumes I-III Cellis, J. E., ed. (1994); "Nucleic Acid Hybridization" Hames, B. D., and Higgins S. J., eds. (1985); "Transcription and Translation" Hames, B. D., and Higgins S. J., eds. (1984); "Animal Cell Culture" Freshney, R. I., ed. (1986); "Immobilized Cells and Enzymes" IRL Press, (1986); "A Practical Guide to Molecular Cloning" Perbal, B., (1984) and "Methods in Enzymology" Vol. 1-317, Academic Press; "PCR Protocols: A Guide To Methods And Applications", Academic Press, San Diego, Calif. (1990); all of which are incorporated by reference as if fully set forth herein. Other general references are provided throughout this document. The procedures therein are believed to be well known in the art and are provided for the convenience of the reader. All the information contained therein is incorporated herein by reference.
Example 1
Materials and Methods
[0207] Plant Material and Growth Conditions
[0208] Experiments were conducted using WT tomato (Solanum lycopersicum cv. MP-1), isogenic independent transgenic homozygous tomato lines expressing different levels of the Arabidopsis AtHXK1 (35S::AtHXK1) [as previously described by Dai et al. (1999)], isogenic transgenic homozygous lines with antisense suppression of the tomato LeHXK1, 2&3 genes, isogenic transgenic homozygous lines expressing GFP or AtHXK1 under the control of the KST1 promoter, and the ABA-deficient mutant Sitiens (Dai et al., 1999) (S. lycopersicum cv. Ailsa Craig).
[0209] Independent antisense-HXK tomato lines, αHK1 and αHK2, were generated following transformation of MP-1 with an antisense construct of StHXK1 (X94302) expressed under the 35S promoter. The potato StHXK1 shares over 80% sequence identity with LeHXK1, 2&3 and conferred suppression of LeHXK1, 2&3 (FIG. 4A). Arabidopsis (Col.) and tomato (MP-1) lines that express GFP or AtHXK1 specifically in guard cells (GCGFP and GCHXK lines, respectively) were generated following transformation with GFP or AtHXK1 expressed under the KST1 promoter (Muller-Rober et al., 1995). Independent transgenic homozygous lines for each construct were then identified. The tomato plants were grown in a temperature-controlled greenhouse under natural growth conditions and the Arabidopsis plants were grown in a walk-in growth chamber kept at 22° C., with an 8-h light/16-h dark photoperiod.
[0210] Stomatal Measurements
[0211] Stomatal aperture and density are determined using the rapid imprinting technique described by Geisler and Sack (2002). This approach allows to reliably score hundreds of stomata from each experiment, each of which is sampled at the same time. Light-bodied vinylpolysiloxane dental resin (Heraeus-Kulzer, Hanau, Germany) is attached to the abaxial leaf side and then removed as soon as it dries (1 min). The resin epidermal imprints are than covered with nail polish, which removed once it had dried out and serves as a mirror image of the resin imprint. The nail-polish imprints are put on glass cover slips and photographed under bright-field microscope. Stomata images are later analyzed to determine aperture size using the ImageJ software (www.rsb.info.nih.gov/ij/) fit-ellipse tool or any other software that can process and analyze images. A microscopic ruler is used for the size calibration. Additional information can be obtained from the software such as stomata width, length, area, perimeter etc.
[0212] To assess stomatal responses, leaflets are cut and immediately immerse in artificial xylem sap solution (AXS) (Wilkinson and Davies, 1997) containing 100 mM sucrose (Duchefa Biochemie) with or without 20 mM N-acetyl glucosamine (NAG, Sigma-Aldrich), 100 mM or 200 mM glucose (Duchefa Biochemie), 100 mM or 200 mM fructose (Sigma-Aldrich), 100 mM 2-deoxyglucose (Sigma-Aldrich), 10 mM or 100 mM mannose (Sigma-Aldrich), 100 mM sorbitol (Sigma-Aldrich) or 100 mM or 200 mM mannitol (Duchefa Biochemie). The sorbitol and mannitol treatments serve as non-metabolic osmotic controls. Imprints are taken 3 h after immersion and stomatal aperture is analyzed. Different plant species can be used as well as, AXS solutions, treatment solutions and different timings to our decision.
[0213] Gas Exchange Analysis
[0214] Gas exchange measurements are assayed using a Li-6400 portable gas-exchange system (LI-COR). Plants are growing under favorable or stressed conditions, and measurements are conducted on fully expanded leaf, 5th-6th from top in the case of tomato. All measurements are conducted between 10:00 AM and 2:00 PM. We are inducing photosynthesis under saturating light (1000-1200 μmol m-2 sec-1) with 370 μmol mol-1 CO2 surrounding the leaf (Ca). The amount of blue light is set to 15% photosynthetically active photon flux density to optimize stomatal aperture. The leaf-to-air VPD (Vapor pressure deficit) is kept at around 1 to 2.5 kPa and leaf temperature is kept at around 25° C., during all measurements. Once a steady state is reached, measurements are done. It is possible to tune each of the above mentioned parameters. Each measurement contains data of photosynthesis (μmol CO2 m-2 s-1), transpiration (mmol H2O m-2 s-1), Stomatal conductance (mol H2O m-2 s-1), and calculated instantaneous water use efficiency (μmol CO2 mmol-1 H2O). Additional data obtained from each measurement are mesophyll conductance for CO2 (mol CO2 m-2 s-1 bar-1), electron transport rate, calculated from PS (photosystem) II quantum yield and internal CO2 concentrations (Ci).
[0215] For stomatal conductance (gs) measurements the leaf conductance steady-state porometer LI-1600 (LI-COR, Lincoln, Nebr.) is used according to manufacture instructions.
[0216] Whole-Plant Transpiration Measurements
[0217] Whole-plant transpiration rates and relative daily transpiration (RDT) are determined using a wide-screen lysimeter-scale system, which allows measurements of up to 160 plants simultaneously. Plants are planted in 3.9-L pots and grow under controlled conditions. Each pot is placed on a temperature-compensated load cell with digital output and is sealed to prevent evaporation from the surface of the growth medium. A wet vertical wick made of 0.15 m2 cotton fibers partially submerged in a 1-L water tank is placed on a similar load cell and use as a reference for the temporal variations in the potential transpiration rate. The output of the load cells is monitored every 10 s and the average readings over 3 min are logged in a data logger for further analysis. The output data includes whole plant transpiration, plant weight, light intensity, vapor pressure deficit (VPD), temperature, stomatal conductance, water use efficiency and additional environmental and physiological parameters. The whole-plant transpiration rate is calculated by a numerical derivative of the load cell output following a data-smoothing process (Sade et al., 2010). The plant's daily transpiration rate is normalized to the total plant weight and the data for neighboring submerged wick and these figures are averaged for a given line over all plants (amount taken up by the wick daily=100%). Water use efficiency is calculated from the daily weight added against the daily water loss for each plant. Plants RDT is monitored under different growth conditions to our decision: Normal irrigation, drought, salt treatment and more. It is possible to shift growth conditions on a daily bases and to monitor plants responses.
[0218] RNA Extraction, cDNA Generation and Quantitative Real-Time PCR Expression Analysis (Based on Goren 2011, Kandel-Kfir 2006)
[0219] Tissue samples are snap-frozen and homogenize in liquid nitrogen. RNA is extracted using the EZ-RNA kit (Biological Industries, Kibbutz Bet Haemek, Israel), with up to 500 μl of frozen homogenized tissue per extraction tube. At least four independent extractions are performed for each tissue set. The extractions are carried out according to the manufacturer's protocol, including two optional washes in 2 M LiCl. RNA pellets are than suspended in 25 μl DEPC-treated H2O and treated with DNase (Ambion, Austin, Tex., USA) according to the manufacturer's instructions. RNA presence is confirmed by gel electrophoresis and DNA degradation is confirmed by PCR. RNA (≦1 μg) from each sample is than reverse-transcribed to cDNA using MMLV RT (ProMega, Madison, Wis., USA) in a 25-μl reaction, with 2 μl random primers and 1 μl mixed poly-dT primers (18-23 nt). All cDNA samples are diluted 1:8 in DEPC-treated water.
[0220] Real-time reactions are prepared using SYBR Green mix (Eurogentec S.A., Seraing, Belgium) in 10 μl volumes with 4 μl diluted cDNA per reaction, two replicates per cDNA sample. Reactions run in a RotorGene 6000 cycler (Corbett, Mortlake, New South Wales, Australia), 40 cycles per run, with sampling after each cycle. Following an initial pre-heating step at 95° C. for 15 min, there are 40 cycles of amplification consisting of 10 s at 95° C., 15 s at 55° C., 10 s at 60° C. and 20 s at 72° C. Results are than interpreted using RotorGene software, two duplicates per sample. Data are normalized using SlCyP as a reference gene (cyclophilin--accession no. M55019). Primers used for amplification: SlCyP--CGTCGTGTTTGGACAAGTTG (SEQ ID NO: 1) and CCGCAGTCAGCAATAACCA (SEQ ID NO: 2). The primers for SlHXKs (LeHXKs) are as follows: for SlHXK1-GACTTGCTGGGAGAGGAGT (SEQ ID NO: 3) and AAGGTACATTGAATGAGAGGCA (SEQ ID NO: 4); for SlHXK2-GTCCTCCCATCTTCCCTTG (SEQ ID NO: 5) and CCCAAGTACATACCAGAACAT (SEQ ID NO: 6); for SlHXK3-GCGATATTATCACCTCTCGTG (SEQ ID NO: 7) and CTGCTTCTCTCCGTCTTTAAA (SEQ ID NO: 8); and for SlHXK4-GCTGAGGACACCTGATATATG (SEQ ID NO: 9) and GATCGGATTTTACCCCAGCTA (SEQ ID NO: 10).
[0221] Protein Extraction and Analysis of Hexokinase Activity
[0222] Protein extraction from plant leaves is performed with 1 to 2 g of plant tissue homogenized in 4 volumes of extraction buffer (50 mM Hepes, pH 7.6, 1 mM EDTA, 15 mM KCl, 1 mM MgCl2, 1 mM phenylmethylsulfonyl fluoride, 3 mM diethyldithiocabamic acid, and 0.2% PVP). The mixture is centrifuged for 25 min at 16,000 g at 48 C., and the supernatant is brought to 80% ammonium sulfate saturation. After centrifugation, the pellet is resuspended in 0.5 mL of washing buffer (50 mM Hepes, pH 7.5, 1 mM EDTA, and 1 mM DTT), desalted on a G-25 Sephadex column (55×11 mm), and used as a crude enzyme extract for subsequent enzymatic analysis.
[0223] Hexokinase activity is measured by enzyme-linked assay according to Schaffer and Petreikov (1997). The assays contain a total volume of 1 mL of 30 mM Hepes-NaOH, pH 7.5, 2 mM MgCl2, 0.6 mM EDTA, 9 mM KCl, 1 mM NAD, 1 mM ATP, and 1 unit of NAD-dependent glucose-6-phosphate dehydrogenase (G6PDH from Leuconostoc mesenteroides; Sigma). To assay glucose phosphorylation, the reaction is initiated with 2 mM glucose. Reactions are conducted at 37° C., and absorption at 340 nm is monitored continuously. (For additional information see Dai et al. 1999, Schaffer and Petreikov, 1997).
[0224] Monitoring Nitric Oxide Production in Guard Cells
[0225] Detection of nitric oxide (NO) levels in stomata is performed as follows:
[0226] Epidermal peels are prepared and incubated in MES buffer [25 mM MES-KOH, pH=6.15 and 10 mM KCl (MES, 2-(N-morpholino)-ethane sulfonic acid; Sigma-Aldrich] with or without 20 mM NAG, for 2.5 h under steady light, and then loaded with 60 μM NO indicator dye, DAF-2DA (4,5-diaminofluorescein diacetate; Sigma-Aldrich), diluted in MES buffer with or without 20 mM N-acetyl glucosamine (NAG, Sigma-Aldrich) and left for an additional 50 min. Then, the peels are washed with MES 3 times and re-incubated for 30 min in the buffer (control, set as 100% fluorescence) or in 100 mM sorbitol, 100 mM sucrose and 20 mM NAG. The peels are then photographed under a microscope (see Materials and Methods, "Confocal microscopy imaging"). Three to four biological repeats containing 20-30 stomata each are included in each experiment and each experiment is repeated several times. Images are analyzed using the ImageJ software histogram tool to evaluate fluorescence intensity and the fit-ellipse tool to determine stomatal aperture. It is possible to use epidermal strips from different species, use different treatments solutions and different timings, all to our decision.
[0227] Confocal Microscopy Imaging
[0228] Images are acquired using the OLYMPUS IX 81 (Japan) inverted laser scanning confocal microscope (FLUOVIEW 500) equipped with a 488-nm argon ion laser and a 60×1.0 NA PlanApo water immersion objective. Nitric oxide-DAF-2DA (4,5-diaminofluorescein diacetate; Sigma-Aldrich) fluorescence is excited by 488-nm light and the emission is collected using a BA 505-525 filter. GFP is excited by 488-nm light and the emission is collected using a BA 505-525 filter. A BA 660 IF emission filter is used to observe chlorophyll autofluorescence. Confocal optical sections are obtained at 0.5-μm increments. The images are color-coded green for GFP and magenta for chlorophyll autofluorescence.
[0229] Thermal Imaging
[0230] Leaf temperature is a reliable tool for determine transpiration variation among different conditions and different plant species. High temperatures are associated with closed stomata and low transpiration, while low temperature points out for open stomata and high transpiration. For thermal imaging, leaves are imaged using a thermal camera (ThermaCAM model SC655; FLIR Systems). Pictures are later analyzed using the ThermaCAM researcher pro 2.10 software. The experiments are repeated several times. Data are means±SE from five biological repeats per line; four leaves are analyzed per plant.
[0231] Use of KST1 as a Guard Cell Specific Promoter
[0232] The KST1 potassium channel in potato (Solanum tuberosum L.) has been shown to be expressed specifically in guard cells (Muller-Rober et al., 1995). Later, by GUS activity and staining assay it has been demonstrated that KST1 promoter segment can be used to express genes exclusively in guard cells (Plesch et al., 2001). Using this knowledge, transgenic tomato and Arabidopsis plants were generated overexpressing Arabidopsis hexokinase1 (KST::AtHXK1) or GFP (green fluorescence protein) (as a control for exclusive expression) specifically in guard cells in the following procedures:
1. Creation of binary vector containing an insert of AtHXK1 cDNA under KST1 promoter followed by terminator. 2. Creation of binary vector containing an insert of GFP gene under KST1 promoter followed by terminator. 3. Plant transformation. 4. Identification of plants containing KST1::AtHXK1 trait.
[0233] Creation of a Binary Vector Containing an Insert of AtHXK1 cDNA or GFP Under KSTI Promoter Followed by Terminator.
[0234] The binary vector pGreen0029 was used (Hellens et al., 2000b) for transformation into tomato and Arabidopsis plants. The KST1 promoter was ligated upstream the AtHXK1 coding sequence (isolated by (Dai et al., 1995) or GFP followed by a terminator (See FIGS. 17A-B).
Example 2
Sucrose Stimulates Stomatal Closure
[0235] To examine the effect of Suc on stomata, intact wild-type (WT) tomato leaflets were immersed in artificial apoplastic solutions (Wilkinson and Davies, 1997) containing either 100 mM Suc or 100 mM sorbitol, a non-metabolic sugar used as an osmotic control, and measured stomatal aperture. Suc decreased stomatal aperture size by 29% relative to sorbitol (FIGS. 1A, B). Sucrose is a disaccharide that has to be cleaved. It may be cleaved by cell wall (apoplastic) invertases, yielding glucose (Glc) and fructose (Fru) in equal proportions (Granot, 2007) and resulting in additional extracellular osmolarities approaching 200 mOsm/L, as compared to the 100 mOsm/L of the original Suc added. We, therefore, compared the effects of 100 mM sucrose, 100 mM Glc+100 mM Fru and 200 mM Glc or Fru with the effect of 200 mM mannitol, which was used as an additional osmotic control. All of the sugar combinations decreased the size of stomatal apertures, as compared to the effect of 200 mM mannitol (FIG. 1C), supporting an osmotic-independent role for sugars in the regulation of stomatal closure.
Example 3
Sucrose Stimulates Stomatal Closure Via Hexokinase
[0236] Sucrose may be cleaved by either apoplastic (extracellular) invertase or enter the cells via sucrose transporters and then be cleaved by intracellular sucrose-cleaving enzymes to yield the hexoses Glc and Fru. The hexoses Glc and Fru must be phosphorylated by hexose-phosphorylating enzymes (Granot, 2007). In plants, hexokinases (HXK) are the only enzymes that can phosphorylate Glc and may also phosphorylate Fru (Granot, 2007, 2008). HXKs are intracellular enzymes known to play both kinetic and sugar-signaling roles (Rolland et al., 2006). To examine whether Suc stimulates stomatal closure via HXK, the effect of Suc was tested in the presence of N-acetyl glucosamine (NAG), an efficient inhibitor of HXK activity (Hofmann and Roitsch, 2000). NAG almost completely abolished the effect of Suc and prevented stomatal closure, supporting a role for HXK in the regulation of stomatal closure (FIG. 1B).
Example 4
Increased Expression of HXK Enhances Stomatal Closure
[0237] To further explore whether HXK mediates stomatal closure, the effect of Suc was examined on well-characterized transgenic tomato plants expressing the Arabidopsis HXK 1 (AtHXK1) under the control of the global non-specific 35S promoter (Dai et al., 1999). The stomatal aperture of AtHXK1-expressing plants (the HK4 line, which has a level of HXK activity that is 5 times higher than that of WT plants) was reduced by 21% relative to the control plants even under the control conditions (100 mM sorbitol) (FIG. 1B), indicating that increased expression of HXK induces stomatal closure. The addition of Suc caused the stomata to close even further (FIG. 1B) and the HXK inhibitor NAG abolished the closing effect of Suc, further supporting a role for HXK in the regulation of stomatal closure (FIG. 1B).
Example 5
Direct Correlation Between HXK Activity, Stomatal Closure and Reduced Transpiration
[0238] To examine the effect of HXK on tomato stomata, the stomatal apertures and conductance of tomato lines expressing increasing levels of AtHXK1 were measured. (The HK37, HK4 and HK38 lines have levels of HXK activity that are 2, 5 and 6 times higher than those of WT plants, respectively) (Dai et al., 1999). The stomatal densities of the AtHXK1-expressing lines are similar to those of WT plants (Table S1), yet both stomatal aperture and conductance were significantly reduced, in direct correlation with the level of AtHXK1 expression (FIGS. 2A, 2B). Furthermore, continued measurement of transpiration over the course of the day revealed that AtHXK1 lowered the transpiration rate per unit leaf area in the AtHXK1-expressing lines, in correlation with the level of AtHXK1 expression (FIG. 2C), so that the cumulative whole-plant relative daily transpiration per unit leaf area (RDT) was clearly negatively correlated with HXK activity (FIG. 2D).
[0239] To rule out the possibility that the observed decrease in transpiration was the result of inhibitory effects of AtHXK1 on root water uptake or stem water transport, reciprocal grafting experiments were performed. HK4 shoots were grafted onto WT roots and WT shoots were grafted onto HK4 roots (FIG. 3A). Continued measurements of the transpiration rates and cumulative whole-plant relative daily transpiration per unit leaf area of the grafted plants indicated that decreased transpiration was generally associated with HK4 shoots, with the roots having only minor influence (FIGS. 3B, 3C). To further examine the effect of HK4 stems on transpiration, triple-grafted plants were generated in which HK4 interstock replaced a portion of the stem of WT plants (FIG. 3D). The HK4 interstock had no effect on RDT (FIG. 3E), indicating that the decreased transpiration of AtHXK1-expressing plants was the result of reduced transpiration by the leaves and not reduced water uptake by the roots or attenuated transport through the stem. The effect of AtHXK1 on leaf transpiration further indicates that HXK controls stomatal behavior that affects the transpiration of intact whole plants.
Example 6
Suppression of HXK Inhibits Stomatal Closure
[0240] The role of HXK in stomatal closure was further examined using tomato and Arabidopsis plants with antisense suppression and knockdown mutants of HXK, respectively. Four HXKs are known in tomato plants, three of which (LeHXK1, 2 and 3) are mitochondria-associated HXKs similar to the sugar sensor AtHXK1 (Granot, 2007, 2008). Unlike the stomatal closure observed in tomato plants expressing high level of AtHXK1 (FIGS. 2A, B), stomatal closure in tomato lines (αHK1 and αHK2) with antisense suppression of LeHXK1, 2&3 (FIG. 4A) was diminished in response to Suc treatments (FIG. 4B). Similarly, the Arabidopsis AtHXK1-knockout gin2-1 mutant had higher stomatal conductance and a higher transpiration rate, as compared to wild-type control plants (FIGS. 8E, F), supporting the hypothesis that HXK plays a role in the regulation of stomatal closure.
Example 7
HXK Mediates Stomatal Closure Independent of Downstream Metabolism of the Phosphorylated Sugars
[0241] To examine whether downstream metabolism of the phosphorylated sugars is required for stomatal closure, the effects of mannose (a glucose epimer at the second carbon atom) and 2-dexoxyglucose (2-dG--a glucose analog) were tested. Both of these sugars are phosphorylated by HXK, but 2-dG is not further metabolized and mannose is poorly metabolized (Klein and Stitt, 1998; Pego et al., 1999). Both mannose and 2-dG reduced stomatal aperture (FIG. 5). A lower concentration of mannose (10 mM) also reduced stomatal aperture more than 100 mM glucose (FIG. 5), in line with previous observations that mannose is more potent than glucose with regard to HXK-mediated sugar effects (Jang and Sheen, 1994; Pego et al., 1999). Moreover, the closure effect of 10 mM mannose further supports an osmotic-independent role of sugars in the stimulation of stomatal closure. The results with mannose and 2-dG suggest that HXK stimulates stomatal closure independent of downstream metabolism of the phosphorylated sugars.
Example 8
Sucrose Stimulates an ABA-Signaling Pathway in Guard Cells
[0242] It has previously been shown that the sugar-signaling effects of HXK, such as the inhibition of photosynthesis and growth, are mediated by abscisic acid (ABA) [for an updated review see Rolland et al. (2006)], a well-known phytohormone that also induces stomatal closure. Therefore, it was speculated that Suc might modulate guard-cell aperture via the HXK and ABA within guard cells. ABA-signaling in guard cells is mediated by the rapid production of nitric oxide (NO), which is required for ABA-induced stomatal closure and serves as an indicator of stomatal-closure stimuli (Garcia-Mata et al., 2003; Neill et al., 2008). To examine the effect of Suc on the ABA-signaling pathway in guard cells, NO levels were monitored within guard cells in response to applications of Suc. Epidermal peels were incubated with Suc and monitored using the fluorescent NO indicator dye diaminofluorescein diacetate (DAF-2DA). Applications of 100 mM sorbitol had no effect on NO levels in guard cells (FIG. 6A). However, the application of 100 mM Suc resulted in a 3.5-fold increase in guard-cell fluorescence, indicating a rapid increase in NO levels, which was correlated with stomatal closure (FIG. 6A). The guard cells of untreated HK4 (AtHXK)-expressing line) epidermal peels exhibited high NO levels, similar to those of Suc-treated WT epidermal peels (FIG. 6B), and the addition of Suc to the peeled HK4 epidermis led to even more intense fluorescence (FIG. 6B).
[0243] To further examine the involvement of HXK in the production of NO in guard cells, the HXK inhibitor NAG was used with epidermal peels. NAG not only inhibited the effect of Suc and blocked stomatal closure (FIG. 1B), it also prevented the production of NO (FIG. 6C). Washing out NAG with 100 mM Suc led to the resumption of NO production within less than 30 min (FIGS. 6D, E). These results suggest that Suc elicits a guard cell-specific NO response via HXK.
[0244] To verify that ABA is indeed required for the stomatal NO response to Suc, the same experiments were conducted with the ABA-deficient tomato mutant Sitiens, whose stomata are always open (Neill and Horgan, 1985). Unlike what was observed for the WT plants, treating Sitiens epidermal peels with 100 mM Suc did not result in any increase in fluorescence or stomatal closure, indicating that there was no production of NO (FIG. 6F). However, treating Sitiens peels with externally supplied ABA did trigger the production of NO (FIG. 6F) and stomatal closure. These findings indicate that Sitiens's guard cells retain their ability to respond to externally supplied ABA by producing NO and that only the absence of ABA production in the Sitiens mutant prevents Suc-triggered NO production and stomatal closure. This observation confirms that Sitiens stomata do not respond to Suc due to this mutant's ABA deficiency and that ABA is a vital mediator of the stomatal response to Suc.
Example 9
Guard-Cell Specific Expression of ATHXK1 Induces Stomatal Closure and Reduces Transpiration of Tomato and Arabidopsis Plants
[0245] To examine the role of HXK specifically in guard cells, tomato and Arabidopsis plants were generated that express AtHXK1 under the KST1 guard-cell specific promoter (Muller-Rober et al., 1995). The specific expression of the KST1 promoter in tomato and Arabidopsis guard cells was verified by expression of GFP under the KST1 promoter (GCGFP lines, FIGS. 7A-E). Expression of the KST1 promoter was specific to guard cells in all of the examined plant organs and was not detected in organs that do not have stomata, such as roots (FIG. 7E). Guard-cell specific expression was recorded from early seedling development, as observed in the hypocotyls of seedlings (FIG. 7D), through the stages in which leaves are fully expanded (FIGS. 7A-C).
[0246] Unlike the expression of AtHXK1 under the 35S promoter (Dai et al., 1999; Kelly et al., 2012), the expression of AtHXK1 under the guard-cell specific KST1 promoter (GCHXK lines) had almost no negative growth effect (FIGS. 8A, D). Yet, expression of AtHXK1 under the KST1 promoter reduced both stomatal conductance and transpiration in both tomato and Arabidopsis plants (FIGS. 8B, C, E, F). These results strongly support the hypothesized specific role of HXK in guard cells, regulating stomatal closure.
Example 10
GFP Expression Under the Control of the FBPase Promoter is Specific to Mesophyll Cells
[0247] To discriminate between HXK effects in guard cells versus mesophyll cells the present inventors have created transgenic tomato and Arabidopsis plants expressing HXK under a mesophyllpromoter FBPase (Peleg et al., 2007). The specific expression of FBPase promoter was demonstrated with transgenic tomato and Arabidopsis plants expressing GFP under control of this promoter (designated MCGFP, FIG. 9). Several independent homozygous Arabidopsis and tomato lines with high expression of FBPase::AtHXK1 (named MCHXK plants) were identified.
Example 11
Elevated Expression of Hexokinase in Guard Cells Reduces Whole Plant Transpiration and Increases Water Use Efficiency, as Determined Using Gas Exchange Analysis System
[0248] Using the LI-COR gas exchange system the present inventors have analyzed 10 GCHXK independent lines and discovered a striking increase in water use efficiency in those plants (FIGS. 10A-D). Our data clearly shows that while photosynthesis remained unchanged (FIG. 10C), stomatal conductance (indicating stomatal aperture, FIG. 10B) and transpiration (FIG. 10A) were reduced by 20% and 15% respectively, thus improving water use efficiency from 1.36 in WT to 1.78 in GCHXK lines (FIG. 10D).
Example 12
Elevated Expression of Hexokinase in Guard Cells Reduces Whole Plant Transpiration and Increases Water Use Efficiency, as Determined Using Lysimeter Scales System
[0249] To evaluate water use efficiency in GCHXK plants the present inventors used the precise and sensitive lysimeter scales system, which measures plant weight accumulation and total plant water loss during long lasting experiments, and can monitor more than 160 plants simultaneously under varied irrigation treatments (FIGS. 11A-C). Two independent GCHXK transgenic lines (that exhibited high WUE when measured by LI-COR (FIGS. A-D 10)) were analyzed. The present inventors have discovered that relative daily transpiration of these lines was lower than WT throughout the entire experiment (20 days) (FIGS. 11A-C). Plant weight accumulation and growth were not affected. As a result, there was about 20%-30% increase in WUE in GCHXK lines compare to WT plants.
Example 13
Elevated Expression of Hexokinase in Guard Cells Reduces Whole Plant Transpiration Rate and Stomatal Conductance, without any Negative Effect on Growth, Thus Enhancing Water Use Efficiency
[0250] Using lysimeter scales system we further analyzed water saving and WUE in GCHXK plants, which displayed high WUE when measured by LI-COR (FIGS. 10A-D) and by lysimeter (FIGS. 11A-C). Several parameters were monitored. Parameters for water loss: transpiration rate, stomatal conductance (gs); parameters for growth: total plant weight, total plant leaf area and environmental parameters: light intensity, vapor pressure deficit (VPD). It was found that along the day, the transpiration rates normalized to total leaf area were correlated with environmental changes (light intensity and VPD, FIGS. 12E and 12F respectively). Transpiration rates of GCHXK plants were significantly lower compared with those of WT along the day (FIG. 12A). Accordingly, stomatal conductance was found to be reduced as well (FIG. 12B) proving that in GCHXK plants, water are saved and stomata are more closed. Moreover, by measuring total plant leaf area and weight (FIGS. 12C and 12D respectively), the present inventors discovered that even though plants have consumed less water (FIG. 12A) growth was not impaired, and was even improved as in the case of GCHXK 12 line. Saving water without affecting plant growth improves whole plant water use efficiency.
Example 14
Elevated Expression of Hexokinase in Guard Cells Enhances Drought Tolerance
[0251] To monitor plants behavior under stress conditions the lysimeter scales system was used. After irrigation was fully stopped, plants were exposed to drought stress, which gradually increased each day throughout the experiment. Transpiration rates of WT and GCHXK plants were analyzed for nine consecutive days (FIG. 13). During the first 3 days GCHXK plants transpired less than WT, in line with normal conditions behavior (FIGS. 11A-C; 12A-F), indicating stress was only moderate at that time. However, in the following days (4 and 5), a transition between WT and GCHXK transpiration rates was observed (FIG. 13, *) and WT transpiration was steeply dropped compared with GCHXKs, indicating that WT plants are more sensitive to drought. As seen in moderate stress (days 5 and 6) as well as in severe stress conditions (days 7 and 8), GCHXK transpiration is less sensitive to water limitation compare to WT, displaying slower decline in transpiration throughout the experiment. These results indicate that GCHXK plants have better tolerance to water shortage and that under mild-stress conditions these plants can still function normally. Drought tolerance was also detected while monitoring relative daily transpiration (RDT) of WT and GCHXK plants under drought conditions (FIG. 11A). While shifting from irrigated to drought conditions (FIG. 11A, days 10-11, magnified), a steep reduction in transpiration was observed for WT plants (red arrow). However, GCHXK transpiration was only moderately affected when exposed to drought (green arrow), indicating that these plants have better tolerance to drought.
Example 15
Elevated Expression of Hexokinase in Guard Cells Improves Yield Production
[0252] To examine the effect of GCHXK on yield, fruits number of GCHXK plants was monitored. Neither of the lines exhibited reduced yield, even though transpiration of these lines was found to be lower (FIGS. 10-12). On the contrary, in few lines fruit number was even higher than control (FIGS. 14A-B).
Example 16
Elevated Expression of Hexokinase in Guard Cells Improves Yield Production Under Limited Water Supply Conditions
[0253] For a wide-range yield production assay, plants were grown in a controlled semi-commercial greenhouse under four different water stressed-irrigation regimes. Plants were irrigated either 25% above the recommended irrigation amount (125%), the recommended irrigation (100%) and deficit irrigation (75%, 50% irrigation regimes, FIG. 15A). Fruits were collected and cumulative fruit numbers and total fruit weight of each plant were documented (FIGS. 15B-C). As clearly seen, GCHXK on yield was dramatic. Compare to WT, GCHXK plants had significantly higher yield (fruit number and total fruit weight under all irrigation regimes. Yet, deficit irrigation did not alter fruit number per plant but reduced fruit weight. Interestingly, GCHXK fruit weight under fully stressed conditions (50% irrigation) was higher than control plants at 100% irrigation. GCHXK plants have also better tolerance to water limitation. When lowering the irrigation from 100% to 75%, fruit weight of GCHXK plants was reduced by only 16% while that of WT control plants was reduced by 39%. Hence, in addition to more yield under normal (100%) irrigation conditions (FIGS. 14A-B and FIG. 15B), GCHXK plants also have better tolerance (higher yield) to limited water supply. Together with the transpiration results (FIG. 13), these results indicate that specific expression of HXK in guard cells saves water, increases water use efficiency and improves yield production, not only under normal, but also under drought conditions as well.
Example 17
Elevated Expression of Hexokinase in Guard Cells Reduces Whole Plant Transpiration, Induces Stomatal Closure and Increases Water Use Efficiency in Arabidopsis
[0254] Thermal imaging and gas-exchange analysis were used to determine stomatal aperture, transpiration and WUE in Arabidopsis plants expressing HXK specifically in guard cells (GCHXK, FIGS. 16A-F). The present inventors have discovered that in GCHXK plants, stomatal conductance and transpiration (FIGS. 16A and B respectively, FIG. 8E-F) are significantly reduced compare to WT. Additionally, by using thermal imaging technique, it was found that the leaf temperature of GCHXK plants was higher than WT, which indicates that stomata are more closed (FIG. 16F). In addition, while transpiration was reduced, photosynthesis rates (FIGS. 16C), as well as the mesophyll conductance to CO2 (gm, FIG. 16D) were not affected. Moreover, growth was not affected as well (FIG. 8D). Overall, GCHXK plants had higher water use efficiency (FIG. 16E). These results demonstrate that the same transgenic insertion of hexokinase under guard-cell specific promoter used in the case of Tomato (Solanaceae family) is universally applicable while affecting stomata and increases water use efficiency in the case of Arabidopsis (Brassicaceae family) as well, and that this technique could be implemented in other species as well.
[0255] Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.
[0256] All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.
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Sequence CWU
1
1
108120DNAArtificial sequenceSingle strand DNA oligonucleotide 1cgtcgtgttt
ggacaagttg
20219DNAArtificial sequenceSingle strand DNA oligonucleotide 2ccgcagtcag
caataacca
19319DNAArtificial sequenceSingle strand DNA oligonucleotide 3gacttgctgg
gagaggagt
19422DNAArtificial sequenceSingle strand DNA oligonucleotide 4aaggtacatt
gaatgagagg ca
22519DNAArtificial sequenceSingle strand DNA oligonucleotide 5gtcctcccat
cttcccttg
19621DNAArtificial sequenceSingle strand DNA oligonucleotide 6cccaagtaca
taccagaaca t
21721DNAArtificial sequenceSingle strand DNA oligonucleotide 7gcgatattat
cacctctcgt g
21821DNAArtificial sequenceSingle strand DNA oligonucleotide 8ctgcttctct
ccgtctttaa a
21921DNAArtificial sequenceSingle strand DNA oligonucleotide 9gctgaggaca
cctgatatat g
211021DNAArtificial sequenceSingle strand DNA oligonucleotide
10gatcggattt taccccagct a
21112030DNAArabidopsis thaliana 11aagctctcgc ttacgtggtt tctacactgt
ttttgacgaa cccaccaagc tcgagtagat 60cggtattaga tccatcttag gtttctctaa
tttctctcaa ttcactccaa aattttgatt 120atttcttctt tctggcttgt caattttagt
catttgtaat ccttgctttt gcgatcggaa 180tcgtaaaaat ccgatctttc ttttagattc
gttttgtttt tgattccaaa tcggaaaaat 240gggtaaagta gctgttggag cgactgttgt
ttgcacggcg gcggtttgtg cggtggctgt 300tttggttgtt cgacgacgga tgcagagctc
agggaagtgg ggacgtgttt tggctatcct 360caaggccttt gaagaggatt gtgcgactcc
gatctcgaaa ctgagacaag tggctgatgc 420tatgaccgtt gagatgcatg ctggtcttgc
atccgacggt ggtagcaaac tcaagatgct 480tatcagctac gttgataatc ttccttccgg
ggatgaaaag ggtctctttt atgcattgga 540cctagggggg acaaacttcc gtgtcatgcg
tgtgcttctt ggcgggaagc aagagcgtgt 600tgttaaacaa gaattcgaag aagtttcgat
tcctcctcat ttgatgactg gtggttcaga 660tgagttgttc aattttatag ctgaagctct
tgcgaagttt gtcgctacag aatgcgaaga 720ctttcatctt ccagaaggta gacagaggga
attaggtttc actttctcgt ttcctgttaa 780gcagacttct ctgtcctctg gtagtctcat
caaatggaca aaaggctttt ccatcgaaga 840agcagttgga caagatgttg ttggagcact
taataaggct ctggaaagag ttggtcttga 900catgcgaatc gcagcacttg ttaatgatac
cgttggaaca ctagccggtg gtagatacta 960taacccggat gttgttgctg ctgttatttt
aggcactggg acaaacgcag cctatgttga 1020gcgtgcaacc gcgatcccta aatggcatgg
tctgcttcca aaatcaggag aaatggttat 1080aaacatggaa tggggaaact tcaggtcatc
acatcttcca ttaaccgagt ttgatcacac 1140gctggatttc gagagtctga atccaggcga
acagattctt gagaaaatca tttccggtat 1200gtacttggga gagattttgc gaagagttct
tctaaagatg gctgaagatg ctgctttctt 1260tggcgataca gtcccatcta agctgagaat
accattcatc attaggactc ctcacatgtc 1320ggctatgcac aacgacactt ctccagactt
gaagattgtt gggagcaaga ttaaggatat 1380attggaggtc cctacaactt ctctgaaaat
gagaaaagtt gtgatcagtc tctgcaacat 1440catagcaacc cgaggagctc gtctctctgc
tgctggaatc tatggtattc tgaagaaact 1500gggaagagat actactaaag acgaggaggt
gcagaaatcg gttatagcca tggatggtgg 1560attgtttgag cattacactc agtttagtga
gtgtatggag agctcactaa aagagttgct 1620tggagatgaa gcttcaggaa gcgttgaagt
cactcactcc aatgatggat caggcattgg 1680agctgcgctt cttgctgctt ctcactctct
ctaccttgaa gactcttaaa acctacccaa 1740agagcgccat ttttcggtaa tttactgaaa
gcttttcgct atcagaaaac gcctaagcca 1800agttctaagg cgtcataaaa gaaagcattc
catgttttta ctcttcccca agactttctt 1860tgtagcaaat aagtttcctt gggagaaata
tttgttttca tgttcttcaa aaataaaaga 1920ctcagttctt cagattctgg gattttatta
taaccagata tgttgtaaaa actacaaatt 1980caaagctcac ttcactggag ttctgagtat
ataaagattt catttttcct 203012496PRTArabidopsis thaliana 12Met
Gly Lys Val Ala Val Gly Ala Thr Val Val Cys Thr Ala Ala Val 1
5 10 15 Cys Ala Val Ala Val Leu
Val Val Arg Arg Arg Met Gln Ser Ser Gly 20
25 30 Lys Trp Gly Arg Val Leu Ala Ile Leu Lys
Ala Phe Glu Glu Asp Cys 35 40
45 Ala Thr Pro Ile Ser Lys Leu Arg Gln Val Ala Asp Ala Met
Thr Val 50 55 60
Glu Met His Ala Gly Leu Ala Ser Asp Gly Gly Ser Lys Leu Lys Met 65
70 75 80 Leu Ile Ser Tyr Val
Asp Asn Leu Pro Ser Gly Asp Glu Lys Gly Leu 85
90 95 Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn
Phe Arg Val Met Arg Val 100 105
110 Leu Leu Gly Gly Lys Gln Glu Arg Val Val Lys Gln Glu Phe Glu
Glu 115 120 125 Val
Ser Ile Pro Pro His Leu Met Thr Gly Gly Ser Asp Glu Leu Phe 130
135 140 Asn Phe Ile Ala Glu Ala
Leu Ala Lys Phe Val Ala Thr Glu Cys Glu 145 150
155 160 Asp Phe His Leu Pro Glu Gly Arg Gln Arg Glu
Leu Gly Phe Thr Phe 165 170
175 Ser Phe Pro Val Lys Gln Thr Ser Leu Ser Ser Gly Ser Leu Ile Lys
180 185 190 Trp Thr
Lys Gly Phe Ser Ile Glu Glu Ala Val Gly Gln Asp Val Val 195
200 205 Gly Ala Leu Asn Lys Ala Leu
Glu Arg Val Gly Leu Asp Met Arg Ile 210 215
220 Ala Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala
Gly Gly Arg Tyr 225 230 235
240 Tyr Asn Pro Asp Val Val Ala Ala Val Ile Leu Gly Thr Gly Thr Asn
245 250 255 Ala Ala Tyr
Val Glu Arg Ala Thr Ala Ile Pro Lys Trp His Gly Leu 260
265 270 Leu Pro Lys Ser Gly Glu Met Val
Ile Asn Met Glu Trp Gly Asn Phe 275 280
285 Arg Ser Ser His Leu Pro Leu Thr Glu Phe Asp His Thr
Leu Asp Phe 290 295 300
Glu Ser Leu Asn Pro Gly Glu Gln Ile Leu Glu Lys Ile Ile Ser Gly 305
310 315 320 Met Tyr Leu Gly
Glu Ile Leu Arg Arg Val Leu Leu Lys Met Ala Glu 325
330 335 Asp Ala Ala Phe Phe Gly Asp Thr Val
Pro Ser Lys Leu Arg Ile Pro 340 345
350 Phe Ile Ile Arg Thr Pro His Met Ser Ala Met His Asn Asp
Thr Ser 355 360 365
Pro Asp Leu Lys Ile Val Gly Ser Lys Ile Lys Asp Ile Leu Glu Val 370
375 380 Pro Thr Thr Ser Leu
Lys Met Arg Lys Val Val Ile Ser Leu Cys Asn 385 390
395 400 Ile Ile Ala Thr Arg Gly Ala Arg Leu Ser
Ala Ala Gly Ile Tyr Gly 405 410
415 Ile Leu Lys Lys Leu Gly Arg Asp Thr Thr Lys Asp Glu Glu Val
Gln 420 425 430 Lys
Ser Val Ile Ala Met Asp Gly Gly Leu Phe Glu His Tyr Thr Gln 435
440 445 Phe Ser Glu Cys Met Glu
Ser Ser Leu Lys Glu Leu Leu Gly Asp Glu 450 455
460 Ala Ser Gly Ser Val Glu Val Thr His Ser Asn
Asp Gly Ser Gly Ile 465 470 475
480 Gly Ala Ala Leu Leu Ala Ala Ser His Ser Leu Tyr Leu Glu Asp Ser
485 490 495
131815DNAArabidopsis thaliana 13tttccaactt ttttttttat taatttgggc
caactttttt tggtttgaga attgggcgag 60ggagaaagat gggtaaagtg gcagttgcaa
cgacggtagt gtgttcggtg gcggtatgtg 120cggcggcggc gttgatagta cggaggagaa
tgaaaagcgc agggaaatgg gcaagagtga 180tagagatatt gaaagccttt gaagaagatt
gtgcaacgcc aattgccaaa ttgagacaag 240tggctgatgc tatgactgtt gagatgcatg
ctggtcttgc ttctgaaggt ggcagcaagc 300ttaagatgct tattagctac gttgataatc
ttccttctgg ggatgagact ggttttttct 360atgcgttgga tctaggcgga acaaacttcc
gtgttatgcg tgtgcttctt ggtgggaagc 420acgaccgtgt tgttaaacga gaattcaaag
aagaatctat tcctcctcat ttgatgaccg 480ggaagtcaca tgaattattc gattttatcg
ttgatgttct tgccaagttt gtcgctacag 540aaggcgagga ctttcatctc ccacctggta
gacaacggga actaggtttt actttctcat 600ttccggttaa gcagctatct ttatcctctg
gcactctcat caactggacg aagggctttt 660ccattgacga tacagttgat aaagatgttg
ttggagaact tgttaaagct atggaaagag 720ttgggctgga catgcttgtc gcagcgcttg
ttaatgatac cattggaaca cttgcgggtg 780gtagatacac taacccggat gtcgttgtcg
cagttatttt gggcaccggc acaaatgcag 840cctatgtcga acgtgcacat gcaattccca
aatggcatgg tttgctaccc aaatcaggag 900aaatggtgat caacatggaa tggggaaact
tcaggtcatc acatcttcca ttgacagagt 960acgaccactc tctagatgtc gatagtttga
atcctggtga acagattctt gagaaaatca 1020tttccggaat gtatctggga gaaatcttgc
gtagagttct tctgaagatg gctgaagaag 1080ctgccttctt tggcgatatc gtcccaccta
agctgaaaat accattcatc ataaggaccc 1140cgaacatgtc tgctatgcac agtgatactt
ccccggattt gaaggttgta ggaagcaagt 1200taaaagacat attggaggtc cagactagtt
ctctgaagat gaggaaagtt gtgatcagcc 1260tatgtaacat cattgcaagc cgaggagctc
gtttatctgc tgcggggatc tatggaatcc 1320tcaagaaaat aggaagagac gcaacaaaag
atggagaagc tcagaaatct gtgatagcga 1380tggacggtgg gctattcgag cattacactc
agttcagtga gtcgatgaag agttcattga 1440aagagttgct tggagatgaa gtttcagaga
gtgttgaagt gatactgtcg aatgatggtt 1500caggtgttgg agctgcatta cttgctgctt
ctcactctca gtatctcgaa cttgaagatg 1560actctgaaac aagttaattt taaaagcctt
ttttttgtgt ttaaccttct tctttgtttt 1620gccgttaggg tttaaacaaa taaaaaaaag
taagaaggta aaaatgccct tttgggaaat 1680tttatttttg acaattttca ggaacaataa
aacctggatt cttcatcaaa gctctgggaa 1740attcaaacga ccagccaatg ttgtagaact
atacatatat attcgagttc tttctatgaa 1800cgttcatttt ttccc
181514502PRTArabidopsis thaliana 14Met
Gly Lys Val Ala Val Ala Thr Thr Val Val Cys Ser Val Ala Val 1
5 10 15 Cys Ala Ala Ala Ala Leu
Ile Val Arg Arg Arg Met Lys Ser Ala Gly 20
25 30 Lys Trp Ala Arg Val Ile Glu Ile Leu Lys
Ala Phe Glu Glu Asp Cys 35 40
45 Ala Thr Pro Ile Ala Lys Leu Arg Gln Val Ala Asp Ala Met
Thr Val 50 55 60
Glu Met His Ala Gly Leu Ala Ser Glu Gly Gly Ser Lys Leu Lys Met 65
70 75 80 Leu Ile Ser Tyr Val
Asp Asn Leu Pro Ser Gly Asp Glu Thr Gly Phe 85
90 95 Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn
Phe Arg Val Met Arg Val 100 105
110 Leu Leu Gly Gly Lys His Asp Arg Val Val Lys Arg Glu Phe Lys
Glu 115 120 125 Glu
Ser Ile Pro Pro His Leu Met Thr Gly Lys Ser His Glu Leu Phe 130
135 140 Asp Phe Ile Val Asp Val
Leu Ala Lys Phe Val Ala Thr Glu Gly Glu 145 150
155 160 Asp Phe His Leu Pro Pro Gly Arg Gln Arg Glu
Leu Gly Phe Thr Phe 165 170
175 Ser Phe Pro Val Lys Gln Leu Ser Leu Ser Ser Gly Thr Leu Ile Asn
180 185 190 Trp Thr
Lys Gly Phe Ser Ile Asp Asp Thr Val Asp Lys Asp Val Val 195
200 205 Gly Glu Leu Val Lys Ala Met
Glu Arg Val Gly Leu Asp Met Leu Val 210 215
220 Ala Ala Leu Val Asn Asp Thr Ile Gly Thr Leu Ala
Gly Gly Arg Tyr 225 230 235
240 Thr Asn Pro Asp Val Val Val Ala Val Ile Leu Gly Thr Gly Thr Asn
245 250 255 Ala Ala Tyr
Val Glu Arg Ala His Ala Ile Pro Lys Trp His Gly Leu 260
265 270 Leu Pro Lys Ser Gly Glu Met Val
Ile Asn Met Glu Trp Gly Asn Phe 275 280
285 Arg Ser Ser His Leu Pro Leu Thr Glu Tyr Asp His Ser
Leu Asp Val 290 295 300
Asp Ser Leu Asn Pro Gly Glu Gln Ile Leu Glu Lys Ile Ile Ser Gly 305
310 315 320 Met Tyr Leu Gly
Glu Ile Leu Arg Arg Val Leu Leu Lys Met Ala Glu 325
330 335 Glu Ala Ala Phe Phe Gly Asp Ile Val
Pro Pro Lys Leu Lys Ile Pro 340 345
350 Phe Ile Ile Arg Thr Pro Asn Met Ser Ala Met His Ser Asp
Thr Ser 355 360 365
Pro Asp Leu Lys Val Val Gly Ser Lys Leu Lys Asp Ile Leu Glu Val 370
375 380 Gln Thr Ser Ser Leu
Lys Met Arg Lys Val Val Ile Ser Leu Cys Asn 385 390
395 400 Ile Ile Ala Ser Arg Gly Ala Arg Leu Ser
Ala Ala Gly Ile Tyr Gly 405 410
415 Ile Leu Lys Lys Ile Gly Arg Asp Ala Thr Lys Asp Gly Glu Ala
Gln 420 425 430 Lys
Ser Val Ile Ala Met Asp Gly Gly Leu Phe Glu His Tyr Thr Gln 435
440 445 Phe Ser Glu Ser Met Lys
Ser Ser Leu Lys Glu Leu Leu Gly Asp Glu 450 455
460 Val Ser Glu Ser Val Glu Val Ile Leu Ser Asn
Asp Gly Ser Gly Val 465 470 475
480 Gly Ala Ala Leu Leu Ala Ala Ser His Ser Gln Tyr Leu Glu Leu Glu
485 490 495 Asp Asp
Ser Glu Thr Ser 500 151653DNAArabidopsis thaliana
15tcttcgttct tacaaaacag aaccaaactt cgacaatgtc actcatgttt tcttcccctg
60tcgtcacccc agcactcgga tctttcacct tctcatctcg accacgctcc aattacatcg
120tgatgtccgc cgtccgatct aactctgctt cgacgtgtcc tatactgacc aagtttcaga
180aagactgcgc cactcctaca ccgtacctac gcaacgtagc caacgccatt gctgatgaca
240tgcgagatgg tctagctgtt gaaggaggag gagatctcga gatgatcttg acttttgttg
300acgctttgcc ttctgggaat gaggaagggt tgttctatgc attggattta ggaggtacaa
360attttcgggt gcgtagcgtg caattaggag gaaagaaaga gcgagtctta gctaccgaat
420ctgaacaaat atctatttct caaaagctta tgattggtac aagtgaggag cttttcgggt
480tcattgcttc aaagcttgca aattttgttg caaaggagaa gccaggtcgg tttcttttag
540aagaagggag gaaaagggag ttagggttta ccttttcatt ccctgtgaag caaacctcta
600ttgattcagg cacattaagc aagtggacta aaggctttaa agtgtctgga atggaaggaa
660aaaatgtggt tgcttgttta aatgaagcta tggaagcaca tggactcgat atgcgagttt
720ctgctcttgt aaatgatgga gtgggaacat tagctggagc aaggtattgg gatgaggatg
780tgatggtcgg tgtgattctt ggcactggga ccaatgcttg ttatgtagaa cagaaacatg
840caattcctaa actccgaagc aaatcttctt ctggaacaac gatcataaac actgagtggg
900gagggttctc taagattctt ccgcaaacca tttttgacct agagatggat gagacaagcc
960tgaatcctgg tgaacattta tatgagaaga tgatctcagg gatgtacctt ggtgaaattg
1020taaggagggt tttgctccat atgtgtgaaa ctagtgactt gtttggacac ttcgctcctg
1080ccaaactctc cactcctttg gcactcagga ccgagcatct atgcaaaatg caagaggaca
1140atacagatga tcttcgggat gttggatcaa tcctatacga ctttttagat gtagaggcga
1200atatgaatgc aaggaggaga gtggtggaag tgtgtgacac agtagtgaaa cgcggagggc
1260gtctagcagg agctggtata gtggcaattc tggagaagat tgaaaaagat accaaaagaa
1320tgggttcagg taaaagaacc gttgtggcta tggacggtgc actgtatgag aagtacccac
1380aatatcgaca gtatatgcaa gacgcactag tcgagcttct tggccataag cttgcaagtc
1440acgttgcgat caaacatacc aaagacgtgt ctgggctcgg tgctgctctt ttggcggcca
1500ctaactccat ttactagtac ttggacctct acttcatgag tataggagga cttttggtcc
1560atttgtttgt gtacatttat ataagatatc atatcaatat caataagatt gtaaagagga
1620gcaacgaact agcagaagat tatgtcttgg acc
165316493PRTArabidopsis thaliana 16Met Ser Leu Met Phe Ser Ser Pro Val
Val Thr Pro Ala Leu Gly Ser 1 5 10
15 Phe Thr Phe Ser Ser Arg Pro Arg Ser Asn Tyr Ile Val Met
Ser Ala 20 25 30
Val Arg Ser Asn Ser Ala Ser Thr Cys Pro Ile Leu Thr Lys Phe Gln
35 40 45 Lys Asp Cys Ala
Thr Pro Thr Pro Tyr Leu Arg Asn Val Ala Asn Ala 50
55 60 Ile Ala Asp Asp Met Arg Asp Gly
Leu Ala Val Glu Gly Gly Gly Asp 65 70
75 80 Leu Glu Met Ile Leu Thr Phe Val Asp Ala Leu Pro
Ser Gly Asn Glu 85 90
95 Glu Gly Leu Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val
100 105 110 Arg Ser Val
Gln Leu Gly Gly Lys Lys Glu Arg Val Leu Ala Thr Glu 115
120 125 Ser Glu Gln Ile Ser Ile Ser Gln
Lys Leu Met Ile Gly Thr Ser Glu 130 135
140 Glu Leu Phe Gly Phe Ile Ala Ser Lys Leu Ala Asn Phe
Val Ala Lys 145 150 155
160 Glu Lys Pro Gly Arg Phe Leu Leu Glu Glu Gly Arg Lys Arg Glu Leu
165 170 175 Gly Phe Thr Phe
Ser Phe Pro Val Lys Gln Thr Ser Ile Asp Ser Gly 180
185 190 Thr Leu Ser Lys Trp Thr Lys Gly Phe
Lys Val Ser Gly Met Glu Gly 195 200
205 Lys Asn Val Val Ala Cys Leu Asn Glu Ala Met Glu Ala His
Gly Leu 210 215 220
Asp Met Arg Val Ser Ala Leu Val Asn Asp Gly Val Gly Thr Leu Ala 225
230 235 240 Gly Ala Arg Tyr Trp
Asp Glu Asp Val Met Val Gly Val Ile Leu Gly 245
250 255 Thr Gly Thr Asn Ala Cys Tyr Val Glu Gln
Lys His Ala Ile Pro Lys 260 265
270 Leu Arg Ser Lys Ser Ser Ser Gly Thr Thr Ile Ile Asn Thr Glu
Trp 275 280 285 Gly
Gly Phe Ser Lys Ile Leu Pro Gln Thr Ile Phe Asp Leu Glu Met 290
295 300 Asp Glu Thr Ser Leu Asn
Pro Gly Glu His Leu Tyr Glu Lys Met Ile 305 310
315 320 Ser Gly Met Tyr Leu Gly Glu Ile Val Arg Arg
Val Leu Leu His Met 325 330
335 Cys Glu Thr Ser Asp Leu Phe Gly His Phe Ala Pro Ala Lys Leu Ser
340 345 350 Thr Pro
Leu Ala Leu Arg Thr Glu His Leu Cys Lys Met Gln Glu Asp 355
360 365 Asn Thr Asp Asp Leu Arg Asp
Val Gly Ser Ile Leu Tyr Asp Phe Leu 370 375
380 Asp Val Glu Ala Asn Met Asn Ala Arg Arg Arg Val
Val Glu Val Cys 385 390 395
400 Asp Thr Val Val Lys Arg Gly Gly Arg Leu Ala Gly Ala Gly Ile Val
405 410 415 Ala Ile Leu
Glu Lys Ile Glu Lys Asp Thr Lys Arg Met Gly Ser Gly 420
425 430 Lys Arg Thr Val Val Ala Met Asp
Gly Ala Leu Tyr Glu Lys Tyr Pro 435 440
445 Gln Tyr Arg Gln Tyr Met Gln Asp Ala Leu Val Glu Leu
Leu Gly His 450 455 460
Lys Leu Ala Ser His Val Ala Ile Lys His Thr Lys Asp Val Ser Gly 465
470 475 480 Leu Gly Ala Ala
Leu Leu Ala Ala Thr Asn Ser Ile Tyr 485
490 172074DNAArabidopsis thaliana 17aatccataaa tctcttcctc
tctctctttc ttatttaatc gtcacttgac cgccggtaag 60aaactcgaga gtttgaatcg
ttttttaacc ctcgccggtg attattgact ggaaaacacc 120aaacgaatcg aagcagttag
tttacaggta gagcagtgac agctgagcaa aaaaaaaggc 180ggcggaaaaa aaagtagtag
tagttgatga tgatgaagat ggtgaaacag attcgtatag 240ggttttctaa tcggcggtgt
tcgctgttcg tgaactcttt attggagtaa gtaaaaagcg 300ccagagagag agatattgtg
tatttgtctt tttggttctg ttttgggatg gggaaagtgg 360cggttgcgtt tgcggcggtt
gctgttgttg cggcttgttc tgttgccgcg gtgatggttg 420ggaggaggat gaagagtcgg
aggaaatgga ggactgttgt tgagattttg aaagagttgg 480aggatgattg tgatactccg
gttgggaggt tgaggcaagt ggttgatgct atggccgtgg 540agatgcacgc tggtttggct
tctgaaggtg gctctaagct taaaatgctc ctcactttcg 600tcgatgattt gcccactggg
agggagaaag gtacttatta tgcacttcac cttggaggca 660cttattttag gattttaagg
gttcttctgg gtgatcaaag gtcttatcta gatgttcaag 720atgttgaacg acacccaata
ccttcacatt tgatgaatag caccagcgag gttcttttca 780actttctcgc cttttccttg
gaaaggttta ttgaaaagga ggaaaacggg tccgattcac 840aaggtgttag aagggaactt
gcatttacgt tctcattccc tgtcaagcat acttctattt 900cttcaggagt tctaattaaa
tggaccaaag gttttgagat tagtgaaatg gttgggcaag 960atatagctga atgtctacaa
ggagctctga acagaagagg cctagatatg catgttgcgg 1020ctcttgtgaa tgatactgtt
ggagccttgt cgcttggata ttatcacgat ccagatacgg 1080ttgttgcggt tgtatttgga
acaggtagta atgcatgtta cttggaaaga accgatgcca 1140taatcaagtg tcagggtctg
cttacaactt ctggaagcat ggtggtaaat atggagtggg 1200gaaatttttg gtcctctcat
ttgcctagaa cttcgtatga cattgacttg gatgcagaga 1260gttcaaatgc aaatgatatg
ggatttgaga agatgatatc aggaatgtat ctgggtgaca 1320ttgttcgtag agtaattctc
cgcatgtcag aagattctga tatctttgga cccatctcgc 1380ccgtgttatc tgagccttac
gttctaagaa caaattcagt ctcagccata catgaagatg 1440acacacctga gttacaagaa
gtagcaagaa tcttgaaaga cataggggta tcagatgtac 1500cactgaaggt gagaaaacta
gtggtgaaaa tatgcgatgt ggttacacga agagcaggga 1560ggcttgcagc agcaggaata
gcaggaatct tgaagaagat aggccgagat ggaagcgggg 1620gaatcacgag cgggagaagc
agaagtgaaa tccaaatgca gaaaagaaca gttgttgcgg 1680tagaaggagg tttgtacatg
aattacacca tgtttaggga atacatggaa gaagctctcg 1740tagagatact aggagaagaa
gtgagtcaat acgtggtggt taaagccatg gaagatggtt 1800ctagcattgg ctctgctctc
ctcgttgcct ctttacagtc atgaatcatc atcgtcgtcg 1860ttcgtattaa atggtgatat
aattggttat gttcgtcctt gtgtgtgtat atagattatt 1920caatatttgt ttgttgttaa
ttataattat tattattcta tatagctttt gtgtaaatgc 1980ttaggaaggt tttatgtatt
ctttgtttct tctattcgta tgatgtgtac tggaaaatag 2040gtactgggtt tttaagaaaa
tgatttacac tgtc 207418498PRTArabidopsis
thaliana 18Met Gly Lys Val Ala Val Ala Phe Ala Ala Val Ala Val Val Ala
Ala 1 5 10 15 Cys
Ser Val Ala Ala Val Met Val Gly Arg Arg Met Lys Ser Arg Arg
20 25 30 Lys Trp Arg Thr Val
Val Glu Ile Leu Lys Glu Leu Glu Asp Asp Cys 35
40 45 Asp Thr Pro Val Gly Arg Leu Arg Gln
Val Val Asp Ala Met Ala Val 50 55
60 Glu Met His Ala Gly Leu Ala Ser Glu Gly Gly Ser Lys
Leu Lys Met 65 70 75
80 Leu Leu Thr Phe Val Asp Asp Leu Pro Thr Gly Arg Glu Lys Gly Thr
85 90 95 Tyr Tyr Ala Leu
His Leu Gly Gly Thr Tyr Phe Arg Ile Leu Arg Val 100
105 110 Leu Leu Gly Asp Gln Arg Ser Tyr Leu
Asp Val Gln Asp Val Glu Arg 115 120
125 His Pro Ile Pro Ser His Leu Met Asn Ser Thr Ser Glu Val
Leu Phe 130 135 140
Asn Phe Leu Ala Phe Ser Leu Glu Arg Phe Ile Glu Lys Glu Glu Asn 145
150 155 160 Gly Ser Asp Ser Gln
Gly Val Arg Arg Glu Leu Ala Phe Thr Phe Ser 165
170 175 Phe Pro Val Lys His Thr Ser Ile Ser Ser
Gly Val Leu Ile Lys Trp 180 185
190 Thr Lys Gly Phe Glu Ile Ser Glu Met Val Gly Gln Asp Ile Ala
Glu 195 200 205 Cys
Leu Gln Gly Ala Leu Asn Arg Arg Gly Leu Asp Met His Val Ala 210
215 220 Ala Leu Val Asn Asp Thr
Val Gly Ala Leu Ser Leu Gly Tyr Tyr His 225 230
235 240 Asp Pro Asp Thr Val Val Ala Val Val Phe Gly
Thr Gly Ser Asn Ala 245 250
255 Cys Tyr Leu Glu Arg Thr Asp Ala Ile Ile Lys Cys Gln Gly Leu Leu
260 265 270 Thr Thr
Ser Gly Ser Met Val Val Asn Met Glu Trp Gly Asn Phe Trp 275
280 285 Ser Ser His Leu Pro Arg Thr
Ser Tyr Asp Ile Asp Leu Asp Ala Glu 290 295
300 Ser Ser Asn Ala Asn Asp Met Gly Phe Glu Lys Met
Ile Ser Gly Met 305 310 315
320 Tyr Leu Gly Asp Ile Val Arg Arg Val Ile Leu Arg Met Ser Glu Asp
325 330 335 Ser Asp Ile
Phe Gly Pro Ile Ser Pro Val Leu Ser Glu Pro Tyr Val 340
345 350 Leu Arg Thr Asn Ser Val Ser Ala
Ile His Glu Asp Asp Thr Pro Glu 355 360
365 Leu Gln Glu Val Ala Arg Ile Leu Lys Asp Ile Gly Val
Ser Asp Val 370 375 380
Pro Leu Lys Val Arg Lys Leu Val Val Lys Ile Cys Asp Val Val Thr 385
390 395 400 Arg Arg Ala Gly
Arg Leu Ala Ala Ala Gly Ile Ala Gly Ile Leu Lys 405
410 415 Lys Ile Gly Arg Asp Gly Ser Gly Gly
Ile Thr Ser Gly Arg Ser Arg 420 425
430 Ser Glu Ile Gln Met Gln Lys Arg Thr Val Val Ala Val Glu
Gly Gly 435 440 445
Leu Tyr Met Asn Tyr Thr Met Phe Arg Glu Tyr Met Glu Glu Ala Leu 450
455 460 Val Glu Ile Leu Gly
Glu Glu Val Ser Gln Tyr Val Val Val Lys Ala 465 470
475 480 Met Glu Asp Gly Ser Ser Ile Gly Ser Ala
Leu Leu Val Ala Ser Leu 485 490
495 Gln Ser 192039DNAArabidopsis thaliana 19gcgtccttct
tctgactctt accaaaattt tatcaatctc tccctcgccg cgccttttcg 60ccggaaggac
ataactttta ccgggatttg ttgtctacaa gggaacctcg agcgcaaggc 120tgtttaatcg
gaagaatctg gagtcacgga gaagcaaaaa tgaacaaaaa gagagaccct 180ttatgctgag
acggtttaat tttgtatctg tggttataga aaggaatcgt ttgcgatgaa 240atggagcctt
atagggttat tctgatcggt ggttgaagcg ttgtcatttg catttcgtgt 300cattctggtt
atagactctg tttcaattgc ttctcgtcgt aaaggcgttt gcttttgtga 360gtaatttatc
tcgattcttc tgaaagacgt agaaagagag aaggttgtgt atagatattg 420gaaatgggga
aggttttggt gatgttgacg gcagctgcgg ctgtggtggc ttgttcagtg 480gcgactgtga
tggtgagaag gaggatgaaa gggaggagga aatggaggag ggtggtgggt 540ttacttaagg
atttggagga agcttgtgag acgcctttag gaaggttgag gcagatggtt 600gatgccatag
ctgtggagat gcaagctggt ttggtttctg aaggagggtc aaagcttaaa 660atgttgctca
cttttgttga tgatcttccc aatgggagcg agacaggaac ctattatgca 720cttcatcttg
gaggctccta ctttaggata ataaaggttc atctaggtgg tcaaagatca 780tctcttgaag
ttcaagatgt tgaacgacat tccataccaa catctttgat gaatagcact 840agcgaggttc
tcttcgactt tctcgcatca tccttgcaga ggtttattga aaaagaaggg 900aacgatttca
gtttgtcaca acctttaaaa agggaacttg cgtttacttt ttctttccca 960gtcaagcaga
catccatctc atcaggagtt ctaattaaat ggaccaaagg ttttgcaatt 1020agtgaaatgg
ctggggaaga cattgctgaa tgtctacaag gagcgttgaa caagagaggg 1080ctagatattc
gcgttgcagc tcttgtgaat gatactgttg gggctttatc ctttggacat 1140tttcatgacc
cagacacaat tgctgctgtt gtctttggaa caggtagtaa tgcatgttac 1200cttgaacgaa
ctgatgccat aatcaagtgt caaaatccac gcacgacttc tggaagcatg 1260gtggtcaata
tggagtgggg aaacttttgg tcatctcgtc tgccaagaac ttcatatgac 1320cttgagttgg
atgcagagag tatgaattca aatgacatgg gctttgagaa gatgatagga 1380gggatgtatc
tgggcgacat tgtccgcaga gtaattcttc gaatgtcaca agagtccgac 1440atctttggac
ctatctcatc cattttatcc acgcctttcg ttctgagaac aaattctgtc 1500tcagcaatgc
atgaagatga cacatccgag ttacaagaag tagcacgaat cttgaaagat 1560ttaggggtgt
cagaggtacc aatgaaggtg aggaaacttg tagtgaagat ctgcgatgta 1620gtgacacgca
gagcagctag gctagcagca gcaggaattg caggaatctt gaagaaggta 1680gggagagatg
ggagcggagg aggaaggaga agcgataagc agataatgag aagaacagtg 1740gtggcagttg
aaggaggtct gtatttgaac tacaggatgt tcagagaata tatggacgaa 1800gctctgagag
atatactggg agaagatgtg gctcaacacg tagtggtgaa ggccatggaa 1860gatggttcca
gcattggctc tgcattgttg ctggcttcgt cacaaagtgt tcaaacaata 1920ccatccgtat
gaataatttg tgtacatatt tacaatgtat atagccaatt gttttagaag 1980catgtaggtt
tgtgtaaata ttgtaggcac tttttagttt cttctactca tgattaagt
203920502PRTArabidopsis thaliana 20Met Gly Lys Val Leu Val Met Leu Thr
Ala Ala Ala Ala Val Val Ala 1 5 10
15 Cys Ser Val Ala Thr Val Met Val Arg Arg Arg Met Lys Gly
Arg Arg 20 25 30
Lys Trp Arg Arg Val Val Gly Leu Leu Lys Asp Leu Glu Glu Ala Cys
35 40 45 Glu Thr Pro Leu
Gly Arg Leu Arg Gln Met Val Asp Ala Ile Ala Val 50
55 60 Glu Met Gln Ala Gly Leu Val Ser
Glu Gly Gly Ser Lys Leu Lys Met 65 70
75 80 Leu Leu Thr Phe Val Asp Asp Leu Pro Asn Gly Ser
Glu Thr Gly Thr 85 90
95 Tyr Tyr Ala Leu His Leu Gly Gly Ser Tyr Phe Arg Ile Ile Lys Val
100 105 110 His Leu Gly
Gly Gln Arg Ser Ser Leu Glu Val Gln Asp Val Glu Arg 115
120 125 His Ser Ile Pro Thr Ser Leu Met
Asn Ser Thr Ser Glu Val Leu Phe 130 135
140 Asp Phe Leu Ala Ser Ser Leu Gln Arg Phe Ile Glu Lys
Glu Gly Asn 145 150 155
160 Asp Phe Ser Leu Ser Gln Pro Leu Lys Arg Glu Leu Ala Phe Thr Phe
165 170 175 Ser Phe Pro Val
Lys Gln Thr Ser Ile Ser Ser Gly Val Leu Ile Lys 180
185 190 Trp Thr Lys Gly Phe Ala Ile Ser Glu
Met Ala Gly Glu Asp Ile Ala 195 200
205 Glu Cys Leu Gln Gly Ala Leu Asn Lys Arg Gly Leu Asp Ile
Arg Val 210 215 220
Ala Ala Leu Val Asn Asp Thr Val Gly Ala Leu Ser Phe Gly His Phe 225
230 235 240 His Asp Pro Asp Thr
Ile Ala Ala Val Val Phe Gly Thr Gly Ser Asn 245
250 255 Ala Cys Tyr Leu Glu Arg Thr Asp Ala Ile
Ile Lys Cys Gln Asn Pro 260 265
270 Arg Thr Thr Ser Gly Ser Met Val Val Asn Met Glu Trp Gly Asn
Phe 275 280 285 Trp
Ser Ser Arg Leu Pro Arg Thr Ser Tyr Asp Leu Glu Leu Asp Ala 290
295 300 Glu Ser Met Asn Ser Asn
Asp Met Gly Phe Glu Lys Met Ile Gly Gly 305 310
315 320 Met Tyr Leu Gly Asp Ile Val Arg Arg Val Ile
Leu Arg Met Ser Gln 325 330
335 Glu Ser Asp Ile Phe Gly Pro Ile Ser Ser Ile Leu Ser Thr Pro Phe
340 345 350 Val Leu
Arg Thr Asn Ser Val Ser Ala Met His Glu Asp Asp Thr Ser 355
360 365 Glu Leu Gln Glu Val Ala Arg
Ile Leu Lys Asp Leu Gly Val Ser Glu 370 375
380 Val Pro Met Lys Val Arg Lys Leu Val Val Lys Ile
Cys Asp Val Val 385 390 395
400 Thr Arg Arg Ala Ala Arg Leu Ala Ala Ala Gly Ile Ala Gly Ile Leu
405 410 415 Lys Lys Val
Gly Arg Asp Gly Ser Gly Gly Gly Arg Arg Ser Asp Lys 420
425 430 Gln Ile Met Arg Arg Thr Val Val
Ala Val Glu Gly Gly Leu Tyr Leu 435 440
445 Asn Tyr Arg Met Phe Arg Glu Tyr Met Asp Glu Ala Leu
Arg Asp Ile 450 455 460
Leu Gly Glu Asp Val Ala Gln His Val Val Val Lys Ala Met Glu Asp 465
470 475 480 Gly Ser Ser Ile
Gly Ser Ala Leu Leu Leu Ala Ser Ser Gln Ser Val 485
490 495 Gln Thr Ile Pro Ser Val
500 211482DNAArabidopsis thaliana 21atgaccagga aagaggtggt
tctggccgtg acggctgcaa ccattacggc ggttgcagca 60ggtgtactaa tgggtcggtg
gatccggagg aaagagcggc ggttgaaaca tacgcagaga 120attttgagga aattcgctag
agaatgcgcc acgccggttt cgaagctttg ggcggtggcg 180gacgccttgg tcgccgacat
gaccgcctct ttaaccgccg agtgttgcgg ttccctcaac 240atgctcgttt cattcaccgg
ttctctccct tccggtgatg agaaaggggt acactatgga 300gtcaacttga gaggcaagga
actattactg ttacgtggga cgctaggtgg taacgaagag 360cctatttccg atgtacagaa
gcatgagatt ccgatccctg acgatgtttt aaatggttct 420ttcaaggagt tgtgcgattt
catatcattg gagcttgtta aatttcttgc gatgaatccc 480ggtggagaag cagaagaagt
gaagaatctc gggtttacgt tgacgcgctc tgttgagcag 540attgggtcac attcaatctc
gtcgatacat aggaagagtt tagcaaatga cgatgatgag 600aaggttttga aagatttggt
gaatgatatg aatgaatcac tggaaacaca cggtctgaaa 660attcggatga acacagcgct
ggtggataat actataggag aattggctgg aggaaggtat 720tatcacaagg acactgtggc
tgcagtatca ttaggtatgg gaaccaacgc tgcttacatt 780gaacaagctc aagagatatc
gaggtggaaa tctgcgatac gtgagccaca agagatcgtt 840gttagcacag agtggggaga
tttcagatct tgccatcttc ctataaccga gttcgatgct 900tctcttgacg cggaaagctt
gaatcccgga catcgaatat ttgagaagat ggtgtcagga 960agatacttag gggagatagt
aagaagagtg ttactaaaaa tgtctgaaga atctgctctc 1020tttggagata cactacctcc
aaaactcaca attccttaca ttttatggtc tccagatatg 1080gctgcaatgc atcaagatat
atccgaagaa cgggagactg taaacaaaaa gctcaaggaa 1140gttttcggta taatggattc
aactcttgcg gcgagagaag ttgtagttga agtatgcgat 1200gtagtcgcgg aacgagcggc
tcgtttagcg ggagcaggaa tagttgggat gataaagaag 1260cttggaagat tagagaagaa
aatgagcatt gtgatagttg aaggaggatt gtatgatcat 1320tatagggtat ttagaaacta
tcttcatagc agcgtttggg aaatgcttgg tgatgagtta 1380tcagatcacg tcgtcattga
gcattctcac ggtggatctg ctgccggagc tctcttcctt 1440gccgcatgtg gcgacggtca
tcaagattct gaaagcaagt ga 148222493PRTArabidopsis
thaliana 22Met Thr Arg Lys Glu Val Val Leu Ala Val Thr Ala Ala Thr Ile
Thr 1 5 10 15 Ala
Val Ala Ala Gly Val Leu Met Gly Arg Trp Ile Arg Arg Lys Glu
20 25 30 Arg Arg Leu Lys His
Thr Gln Arg Ile Leu Arg Lys Phe Ala Arg Glu 35
40 45 Cys Ala Thr Pro Val Ser Lys Leu Trp
Ala Val Ala Asp Ala Leu Val 50 55
60 Ala Asp Met Thr Ala Ser Leu Thr Ala Glu Cys Cys Gly
Ser Leu Asn 65 70 75
80 Met Leu Val Ser Phe Thr Gly Ser Leu Pro Ser Gly Asp Glu Lys Gly
85 90 95 Val His Tyr Gly
Val Asn Leu Arg Gly Lys Glu Leu Leu Leu Leu Arg 100
105 110 Gly Thr Leu Gly Gly Asn Glu Glu Pro
Ile Ser Asp Val Gln Lys His 115 120
125 Glu Ile Pro Ile Pro Asp Asp Val Leu Asn Gly Ser Phe Lys
Glu Leu 130 135 140
Cys Asp Phe Ile Ser Leu Glu Leu Val Lys Phe Leu Ala Met Asn Pro 145
150 155 160 Gly Gly Glu Ala Glu
Glu Val Lys Asn Leu Gly Phe Thr Leu Thr Arg 165
170 175 Ser Val Glu Gln Ile Gly Ser His Ser Ile
Ser Ser Ile His Arg Lys 180 185
190 Ser Leu Ala Asn Asp Asp Asp Glu Lys Val Leu Lys Asp Leu Val
Asn 195 200 205 Asp
Met Asn Glu Ser Leu Glu Thr His Gly Leu Lys Ile Arg Met Asn 210
215 220 Thr Ala Leu Val Asp Asn
Thr Ile Gly Glu Leu Ala Gly Gly Arg Tyr 225 230
235 240 Tyr His Lys Asp Thr Val Ala Ala Val Ser Leu
Gly Met Gly Thr Asn 245 250
255 Ala Ala Tyr Ile Glu Gln Ala Gln Glu Ile Ser Arg Trp Lys Ser Ala
260 265 270 Ile Arg
Glu Pro Gln Glu Ile Val Val Ser Thr Glu Trp Gly Asp Phe 275
280 285 Arg Ser Cys His Leu Pro Ile
Thr Glu Phe Asp Ala Ser Leu Asp Ala 290 295
300 Glu Ser Leu Asn Pro Gly His Arg Ile Phe Glu Lys
Met Val Ser Gly 305 310 315
320 Arg Tyr Leu Gly Glu Ile Val Arg Arg Val Leu Leu Lys Met Ser Glu
325 330 335 Glu Ser Ala
Leu Phe Gly Asp Thr Leu Pro Pro Lys Leu Thr Ile Pro 340
345 350 Tyr Ile Leu Trp Ser Pro Asp Met
Ala Ala Met His Gln Asp Ile Ser 355 360
365 Glu Glu Arg Glu Thr Val Asn Lys Lys Leu Lys Glu Val
Phe Gly Ile 370 375 380
Met Asp Ser Thr Leu Ala Ala Arg Glu Val Val Val Glu Val Cys Asp 385
390 395 400 Val Val Ala Glu
Arg Ala Ala Arg Leu Ala Gly Ala Gly Ile Val Gly 405
410 415 Met Ile Lys Lys Leu Gly Arg Leu Glu
Lys Lys Met Ser Ile Val Ile 420 425
430 Val Glu Gly Gly Leu Tyr Asp His Tyr Arg Val Phe Arg Asn
Tyr Leu 435 440 445
His Ser Ser Val Trp Glu Met Leu Gly Asp Glu Leu Ser Asp His Val 450
455 460 Val Ile Glu His Ser
His Gly Gly Ser Ala Ala Gly Ala Leu Phe Leu 465 470
475 480 Ala Ala Cys Gly Asp Gly His Gln Asp Ser
Glu Ser Lys 485 490
231862DNASolanum lycopersicum 23gaattcggca ccagctttga tccgatctcc
tctctgtaat ttttattatt tctccttaaa 60aaatcacaaa tctttacttt actcatttca
tttttttctc agtggaccaa ctttttgcca 120acctcaaatt ccggcaaaat gaagaaagtg
acggtgggag tcgccgtggt tggtgcagct 180gcggtgtgtg cagtggcggt gttaatagtg
aatcaccgga tgcggaaatc tagtaaatgg 240ggtcgtgcta tggctattct tcgtgaattt
gaagaaaagt gtaagactca agatgcaaag 300cttaagcaag ttgctgatgc tatgactgtt
gagatgcacg ccggacttgc ttctgaaggc 360ggcagtaagc tcaagatgct tatcacttat
gtggataatc tacccactgg tgatgaagct 420ggcgtcttct atgcgttgga tcttggtgga
acaaattttc gagtattgcg agtgcaattg 480ggtggaaaag atggtggtat tattcatcaa
gaatttgctg aggcatcaat tcctccaagt 540ttgatggttg ggacttcaga tgaacttttt
gattatattg cggctgagct tgcaaaattt 600gttgctgcgg aagaggaaaa atttcatcaa
cctcctggta agcagagaga actaggtttc 660accttctcat tcccagtaat gcagacttca
atcaactccg ggaatattat gcggtggaca 720aaaggcttct ctattgatga tgcggttggt
caagatgttg ttggagaact cacaaaagct 780atgaaaagaa aaggtgtcga tatgcgtgtc
tcagctctgg tgaatgatac cgttggaacg 840ttagctggtg gtaaatatac gcaaaaggat
gtagctgttg ctgttatctt aggtacaggg 900accaatgcag cttatgtgga acgggtgcag
gcaattccaa agtggcatgg tcctgtgcca 960aaatctggtg aaatggttat caatatggaa
tggggtaatt ttaggtcatc ccatctgccg 1020ttgacagagt atgatcatgc attggataat
gagagtttaa atcctggtga acagatattt 1080gagaagatga cttctggcat gtacttggga
gaaattttac gcagagttct acttagggtg 1140gctgaagaag ctggcgtttt tggtgatgag
gtccctccaa agctcaagga accatttgtg 1200ttaaggacac ctgatatgtc tgctatgcat
catgacacat cctctgatct gaaagtggtt 1260ggtgaaaagc tgaaggatat tttagagata
tctaatacct ccttgaagac gagaaaacta 1320gtggttgagc tgtgcaatat cgttgccaca
cgtggggcaa gacttgcagc tgcaggtgta 1380ttgggcatct tgaaaaagat gggaagagat
acgcctaagc agggtggttc agaaaggacg 1440gttatagcca tggatggcgg gttgtatgag
cactatacag aatacaggat gtgtttagag 1500aactctttga aggacttgct gggagaggag
ttagcaacga gcatcgtttt tgtgcactcc 1560aatgatggtt ctggcattgg tgctgctctt
cttgctgcct ctcattcaat gtaccttgaa 1620gatcaagatg cttaggagtg tgacaaaagc
catatcttgc tagagggaac tacctttaga 1680ctcctctcta gcgttttcgg tattttctct
cctttatgta ccatcaaaat gaaattaata 1740atatctccag cagatacatt taacttttat
ctctaggaga gttttcctaa gagtgccttc 1800ataaacacac aattttcatg gataaagagt
ccttttacta cctgaaaaaa aaaaaaaaaa 1860aa
186224498PRTSolanum lycopersicum 24Met
Lys Lys Val Thr Val Gly Val Ala Val Val Gly Ala Ala Ala Val 1
5 10 15 Cys Ala Val Ala Val Leu
Ile Val Asn His Arg Met Arg Lys Ser Ser 20
25 30 Lys Trp Gly Arg Ala Met Ala Ile Leu Arg
Glu Phe Glu Glu Lys Cys 35 40
45 Lys Thr Gln Asp Ala Lys Leu Lys Gln Val Ala Asp Ala Met
Thr Val 50 55 60
Glu Met His Ala Gly Leu Ala Ser Glu Gly Gly Ser Lys Leu Lys Met 65
70 75 80 Leu Ile Thr Tyr Val
Asp Asn Leu Pro Thr Gly Asp Glu Ala Gly Val 85
90 95 Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn
Phe Arg Val Leu Arg Val 100 105
110 Gln Leu Gly Gly Lys Asp Gly Gly Ile Ile His Gln Glu Phe Ala
Glu 115 120 125 Ala
Ser Ile Pro Pro Ser Leu Met Val Gly Thr Ser Asp Glu Leu Phe 130
135 140 Asp Tyr Ile Ala Ala Glu
Leu Ala Lys Phe Val Ala Ala Glu Glu Glu 145 150
155 160 Lys Phe His Gln Pro Pro Gly Lys Gln Arg Glu
Leu Gly Phe Thr Phe 165 170
175 Ser Phe Pro Val Met Gln Thr Ser Ile Asn Ser Gly Asn Ile Met Arg
180 185 190 Trp Thr
Lys Gly Phe Ser Ile Asp Asp Ala Val Gly Gln Asp Val Val 195
200 205 Gly Glu Leu Thr Lys Ala Met
Lys Arg Lys Gly Val Asp Met Arg Val 210 215
220 Ser Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala
Gly Gly Lys Tyr 225 230 235
240 Thr Gln Lys Asp Val Ala Val Ala Val Ile Leu Gly Thr Gly Thr Asn
245 250 255 Ala Ala Tyr
Val Glu Arg Val Gln Ala Ile Pro Lys Trp His Gly Pro 260
265 270 Val Pro Lys Ser Gly Glu Met Val
Ile Asn Met Glu Trp Gly Asn Phe 275 280
285 Arg Ser Ser His Leu Pro Leu Thr Glu Tyr Asp His Ala
Leu Asp Asn 290 295 300
Glu Ser Leu Asn Pro Gly Glu Gln Ile Phe Glu Lys Met Thr Ser Gly 305
310 315 320 Met Tyr Leu Gly
Glu Ile Leu Arg Arg Val Leu Leu Arg Val Ala Glu 325
330 335 Glu Ala Gly Val Phe Gly Asp Glu Val
Pro Pro Lys Leu Lys Glu Pro 340 345
350 Phe Val Leu Arg Thr Pro Asp Met Ser Ala Met His His Asp
Thr Ser 355 360 365
Ser Asp Leu Lys Val Val Gly Glu Lys Leu Lys Asp Ile Leu Glu Ile 370
375 380 Ser Asn Thr Ser Leu
Lys Thr Arg Lys Leu Val Val Glu Leu Cys Asn 385 390
395 400 Ile Val Ala Thr Arg Gly Ala Arg Leu Ala
Ala Ala Gly Val Leu Gly 405 410
415 Ile Leu Lys Lys Met Gly Arg Asp Thr Pro Lys Gln Gly Gly Ser
Glu 420 425 430 Arg
Thr Val Ile Ala Met Asp Gly Gly Leu Tyr Glu His Tyr Thr Glu 435
440 445 Tyr Arg Met Cys Leu Glu
Asn Ser Leu Lys Asp Leu Leu Gly Glu Glu 450 455
460 Leu Ala Thr Ser Ile Val Phe Val His Ser Asn
Asp Gly Ser Gly Ile 465 470 475
480 Gly Ala Ala Leu Leu Ala Ala Ser His Ser Met Tyr Leu Glu Asp Gln
485 490 495 Asp Ala
251770DNASolanum lycopersicummisc_feature(1329)..(1329)n is a, c, g, or t
25cttctttttt tcaatgaatt aatcaaatac ataatcttga tctgatctca tttattcata
60aagaaagaac attattatta ctctaataac aatccttaat taatctgtaa ttttatacga
120tttcggatag taaaaggatg aagaaggcga cggtgggtgc ggtggtggta ggtacagcgg
180cggcggtagc tgtggcggcg ctcgtcatgc gccaccgcat gggtaaatcg agcaaatggg
240cacgtgccag ggcaattctg aaggaattcg aggagaaatg tgccacccca gatgccaagc
300tgaagcaagt ggctgatgct atgacggtgg agatgcacgc cggactcgcc tctgaaggag
360gcagcaagct caagatgctt attagctatg tagacaatct ccctactggt gatgaagcag
420gagtctttta tgcattggat cttggtggaa cgaattttcg agtattgcgg gtacagttgg
480ggggaaaaga tggtgggatt atgcatcaag aatttgcgga ggcatcaatt cctccaaatt
540tgatggttgg aacttcagaa gccctttttg actatattgc ggcagaactt gcaaaattcg
600tagatgaaga aggagaaaaa tttcatccac ctcctggtaa gcagagagaa ttaggcttca
660ccttctcgtt cccaataatg cagacttcaa tcaattctgg aactcttatc aggtggacga
720aaggtttctc cattgatgac acggttggca aagatgttgt tgcagaactg acaaaagcaa
780tgcaaaaacg agaaattgat atgagggtgt cagcgcttgt gaatgatact gttggaacat
840tggctggtgg tagattcacg gataaggatg tatccattgc tgtgatatta ggtactggga
900caaatgcwgc atatgtggaa cgkgctcagg caatycccaa atggcacggt cctctgccta
960actctggaga aatggtgatc aatatggaat ggggtaactt taggtcctcc catcttccct
1020tgacacagta tgataatgct atggataccg atagtttaaa tcccggtgaa cagatatttg
1080agaagatatg ttctggtatg tacttgggag aaattttacg cagagttcta cttagaatgg
1140ctaaagaagc tggcattttt ggcgaggaag ttcctccaaa actcaagaat tcattcatat
1200tgaggacacc tgaaatgtct gctatgcatc atgacacatc ctctgatttg agagtggttg
1260gcgacaagtt gaaggatatc ttagagatat ccaatacctc cttgaagaca aggagattag
1320ttgttgagnt gtgtaatatt gttgcaacac gtggcgccag acttgcggca gctggtatct
1380tgggcattat caaaaagatg ggaaaggata caccgaggga aagtggtcca gaaaagattg
1440tggtagccat ggatggcgga ttgtatgagc actatacaga atacagcaag tgcttggaaa
1500acacattggt tgaattgctt ggaaaggaaa tggcaacaag cattgtcttc aagcacgcga
1560atgatggttc tggcattggc gctgcactcc tcgcggcctc caactctgtg tatgttgaag
1620acaagtgaga gtgatcaaaa tgatatttag ctagaggaaa ctccatttac cttttatatt
1680atgttttttc tctagccttt tcttttttgt tcttctaaca attatttcca gatttattat
1740tcctgggaat gccaacatat ttcttctgga
177026496PRTSolanum lycopersicummisc_feature(398)..(398)Xaa can be any
naturally occurring amino acid 26Met Lys Lys Ala Thr Val Gly Ala Val Val
Val Gly Thr Ala Ala Ala 1 5 10
15 Val Ala Val Ala Ala Leu Val Met Arg His Arg Met Gly Lys Ser
Ser 20 25 30 Lys
Trp Ala Arg Ala Arg Ala Ile Leu Lys Glu Phe Glu Glu Lys Cys 35
40 45 Ala Thr Pro Asp Ala Lys
Leu Lys Gln Val Ala Asp Ala Met Thr Val 50 55
60 Glu Met His Ala Gly Leu Ala Ser Glu Gly Gly
Ser Lys Leu Lys Met 65 70 75
80 Leu Ile Ser Tyr Val Asp Asn Leu Pro Thr Gly Asp Glu Ala Gly Val
85 90 95 Phe Tyr
Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Arg Val 100
105 110 Gln Leu Gly Gly Lys Asp Gly
Gly Ile Met His Gln Glu Phe Ala Glu 115 120
125 Ala Ser Ile Pro Pro Asn Leu Met Val Gly Thr Ser
Glu Ala Leu Phe 130 135 140
Asp Tyr Ile Ala Ala Glu Leu Ala Lys Phe Val Asp Glu Glu Gly Glu 145
150 155 160 Lys Phe His
Pro Pro Pro Gly Lys Gln Arg Glu Leu Gly Phe Thr Phe 165
170 175 Ser Phe Pro Ile Met Gln Thr Ser
Ile Asn Ser Gly Thr Leu Ile Arg 180 185
190 Trp Thr Lys Gly Phe Ser Ile Asp Asp Thr Val Gly Lys
Asp Val Val 195 200 205
Ala Glu Leu Thr Lys Ala Met Gln Lys Arg Glu Ile Asp Met Arg Val 210
215 220 Ser Ala Leu Val
Asn Asp Thr Val Gly Thr Leu Ala Gly Gly Arg Phe 225 230
235 240 Thr Asp Lys Asp Val Ser Ile Ala Val
Ile Leu Gly Thr Gly Thr Asn 245 250
255 Ala Ala Tyr Val Glu Arg Ala Gln Ala Ile Pro Lys Trp His
Gly Pro 260 265 270
Leu Pro Asn Ser Gly Glu Met Val Ile Asn Met Glu Trp Gly Asn Phe
275 280 285 Arg Ser Ser His
Leu Pro Leu Thr Gln Tyr Asp Asn Ala Met Asp Thr 290
295 300 Asp Ser Leu Asn Pro Gly Glu Gln
Ile Phe Glu Lys Ile Cys Ser Gly 305 310
315 320 Met Tyr Leu Gly Glu Ile Leu Arg Arg Val Leu Leu
Arg Met Ala Lys 325 330
335 Glu Ala Gly Ile Phe Gly Glu Glu Val Pro Pro Lys Leu Lys Asn Ser
340 345 350 Phe Ile Leu
Arg Thr Pro Glu Met Ser Ala Met His His Asp Thr Ser 355
360 365 Ser Asp Leu Arg Val Val Gly Asp
Lys Leu Lys Asp Ile Leu Glu Ile 370 375
380 Ser Asn Thr Ser Leu Lys Thr Arg Arg Leu Val Val Glu
Xaa Cys Asn 385 390 395
400 Ile Val Ala Thr Arg Gly Ala Arg Leu Ala Ala Ala Gly Ile Leu Gly
405 410 415 Ile Ile Lys Lys
Met Gly Lys Asp Thr Pro Arg Glu Ser Gly Pro Glu 420
425 430 Lys Ile Val Val Ala Met Asp Gly Gly
Leu Tyr Glu His Tyr Thr Glu 435 440
445 Tyr Ser Lys Cys Leu Glu Asn Thr Leu Val Glu Leu Leu Gly
Lys Glu 450 455 460
Met Ala Thr Ser Ile Val Phe Lys His Ala Asn Asp Gly Ser Gly Ile 465
470 475 480 Gly Ala Ala Leu Leu
Ala Ala Ser Asn Ser Val Tyr Val Glu Asp Lys 485
490 495 272166DNASolanum
lycopersicummisc_feature(1897)..(1897)n is a, c, g, or t 27acatacacca
aaaagtttga tttttttcaa gaaagtatgg gaaaattggt tgtaggtgca 60acagttgtgt
gtactgctgc tgtagtatgt ggggtgacag ttttgttaat gaaacatagg 120gtgaagaatt
ctggggagtg gggaaaagtt gaagctttat tgaaagattt tgaggagaag 180tgtgcaactc
cagtggaaaa attaaagcag gtagctgatg ctatgactgt agagatgcaa 240gctggacttg
cttctgaagg tgggagtaag ctcaagatgc ttattagcta tgttgataac 300cttcctactg
gggatgaaaa aggtctgttt tatgcattgg atctaggcgg cacaaacttt 360cgagtgatgc
gtgtacagtt gggtgggaaa gaaaagcgta tagttaaaca tgaagttaaa 420gaagtttcaa
ttccacagaa tgtgatgact ggatcatcat ctgaagtgtt atttgatttt 480attgccacgg
cacttgcaga atttgtagct acagaaggtg atgattttca tcttccacct 540ggtagacaaa
gggagttagg ctttaccttc tctttccctg tgaaacaatt gtcaattgca 600tcaggaactc
ttattaaatg gacaaagggc ttctccatag aagatttggt tgggcaagat 660gtggttggag
aattagcaaa agcaatggaa agggccggcc ttgacgtgcg tgtggctgca 720ttagtaaatg
atactgttgg aacgttagcg ggaggtcggt acaataatcc tgatgtcatt 780gctgcagtaa
ttttgggtac cggaaccaat gcagcatatg ttgaacgggc tcatgcgatt 840cccaaatggc
atggtctgtt gcctaaatcc ggagaaatgg ttatcaacat ggaatggggt 900aatttccgct
catcacatct tccagtaaca gaatatgacc aaaatcttga tattgagagt 960ttaaaccccg
gtgagcagat ttatgaaaag attatttccg ggatgtatct tggagaaatt 1020ttgcgtagag
tattgtgtag aatggctgaa gaagcttcat tattcggtga ttatgtccca 1080tccaaactga
aagttccttt cgtattgagg actccggaca tggctgctat gcatcacgac 1140gagtctgctg
atctcaaggt tgttggaaat aagctgaagg atatcttaga ggtacctaat 1200tctaccttga
aaatgaggaa aatagttgtg gagctgtgcg atattatcac ctctcgtgga 1260gctcgtcttt
ctgcagcagg aattgtgggc atcctcaaga aattgggaag agacactttt 1320aaagacggag
agaagcagag gtctgtcata gctgtggacg gtgcattgtt tgagcattac 1380accaagttca
gaaattgctt gaaggaaact atgaaagagt tactgggaga tgctgcagat 1440agcacagtca
ttgagctttc taatgatggt tcaggcgttg gagctgcact tttggctgcc 1500tcacattccc
aatacacaga tctcgaggaa tcttgatcat ggtcagagtg acacaaccaa 1560aagtgcctgg
tcaagagttc tctacgttta caatcgcccc tcttcttgct aaagggaacc 1620cacttcttac
tttcttctac gagacgattg aaatatttct tgcttccttg tgcactgtgt 1680catgcaagag
tgaactttga agtgagccat agttcaatat accaaaatga gcacatctct 1740cttcagctaa
acaccaatat gctgaccttt tcactcctgg tgcctctaag aaccatcttt 1800tatcccaagt
gttaacttaa atttttccct cactagccag aaaataaagt taggatacaa 1860ataaagttat
tctagttgcc acatttttgt gtaatcnttt cnaagattta ttgttgccaa 1920aagttactta
ccgggaaaca acctctncat ctctttaagg tagggatagg attttcttac 1980tctataccct
ccctagagtc cacttatgag attacatcag atatgttgtt gtatcgttgc 2040caaaagtttt
ggatttgtgg aattacatcg tatatgttgt tgtatggttg ccaaaagtta 2100tagtgctgag
gtagttcaag cttgtttcta tttttctgga tttttaggaa atctctcctc 2160cattca
216628499PRTSolanum lycopersicum 28Met Gly Lys Leu Val Val Gly Ala Thr
Val Val Cys Thr Ala Ala Val 1 5 10
15 Val Cys Gly Val Thr Val Leu Leu Met Lys His Arg Val Lys
Asn Ser 20 25 30
Gly Glu Trp Gly Lys Val Glu Ala Leu Leu Lys Asp Phe Glu Glu Lys
35 40 45 Cys Ala Thr Pro
Val Glu Lys Leu Lys Gln Val Ala Asp Ala Met Thr 50
55 60 Val Glu Met Gln Ala Gly Leu Ala
Ser Glu Gly Gly Ser Lys Leu Lys 65 70
75 80 Met Leu Ile Ser Tyr Val Asp Asn Leu Pro Thr Gly
Asp Glu Lys Gly 85 90
95 Leu Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Met Arg
100 105 110 Val Gln Leu
Gly Gly Lys Glu Lys Arg Ile Val Lys His Glu Val Lys 115
120 125 Glu Val Ser Ile Pro Gln Asn Val
Met Thr Gly Ser Ser Ser Glu Val 130 135
140 Leu Phe Asp Phe Ile Ala Thr Ala Leu Ala Glu Phe Val
Ala Thr Glu 145 150 155
160 Gly Asp Asp Phe His Leu Pro Pro Gly Arg Gln Arg Glu Leu Gly Phe
165 170 175 Thr Phe Ser Phe
Pro Val Lys Gln Leu Ser Ile Ala Ser Gly Thr Leu 180
185 190 Ile Lys Trp Thr Lys Gly Phe Ser Ile
Glu Asp Leu Val Gly Gln Asp 195 200
205 Val Val Gly Glu Leu Ala Lys Ala Met Glu Arg Ala Gly Leu
Asp Val 210 215 220
Arg Val Ala Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala Gly Gly 225
230 235 240 Arg Tyr Asn Asn Pro
Asp Val Ile Ala Ala Val Ile Leu Gly Thr Gly 245
250 255 Thr Asn Ala Ala Tyr Val Glu Arg Ala His
Ala Ile Pro Lys Trp His 260 265
270 Gly Leu Leu Pro Lys Ser Gly Glu Met Val Ile Asn Met Glu Trp
Gly 275 280 285 Asn
Phe Arg Ser Ser His Leu Pro Val Thr Glu Tyr Asp Gln Asn Leu 290
295 300 Asp Ile Glu Ser Leu Asn
Pro Gly Glu Gln Ile Tyr Glu Lys Ile Ile 305 310
315 320 Ser Gly Met Tyr Leu Gly Glu Ile Leu Arg Arg
Val Leu Cys Arg Met 325 330
335 Ala Glu Glu Ala Ser Leu Phe Gly Asp Tyr Val Pro Ser Lys Leu Lys
340 345 350 Val Pro
Phe Val Leu Arg Thr Pro Asp Met Ala Ala Met His His Asp 355
360 365 Glu Ser Ala Asp Leu Lys Val
Val Gly Asn Lys Leu Lys Asp Ile Leu 370 375
380 Glu Val Pro Asn Ser Thr Leu Lys Met Arg Lys Ile
Val Val Glu Leu 385 390 395
400 Cys Asp Ile Ile Thr Ser Arg Gly Ala Arg Leu Ser Ala Ala Gly Ile
405 410 415 Val Gly Ile
Leu Lys Lys Leu Gly Arg Asp Thr Phe Lys Asp Gly Glu 420
425 430 Lys Gln Arg Ser Val Ile Ala Val
Asp Gly Ala Leu Phe Glu His Tyr 435 440
445 Thr Lys Phe Arg Asn Cys Leu Lys Glu Thr Met Lys Glu
Leu Leu Gly 450 455 460
Asp Ala Ala Asp Ser Thr Val Ile Glu Leu Ser Asn Asp Gly Ser Gly 465
470 475 480 Val Gly Ala Ala
Leu Leu Ala Ala Ser His Ser Gln Tyr Thr Asp Leu 485
490 495 Glu Glu Ser 291774DNASolanum
lycopersicum 29ggcacgagag atctcactgt tacctaaaat gtcggtcacc gttagctcgc
cggccgtccg 60atccttccat gtttcacgat cacctcataa aacgatctct aggccacgtg
tcattatatc 120tgccgtccga tctactgaca gcttaggagt agcaccaatt ttgacgaagt
tgcagaaaga 180ttgtgctact cctcttccag ttttgcgcca cgtggcggat gcaatggccg
atgatatgag 240ggccgggctt gccgtcgacg gcggcagtga tctgaagatg atccttagtt
atgtcgacac 300tctaccaact gggaatgaga aaggcttgtt ttatgcgttg gaccttggtg
gtacaaattt 360ccgggtgcta agggtgcagc taggtggtaa agaagagcgt gtagtcgcca
ctgagtttga 420gcaagtctct ataccccaag aactgatgtt tgctacctcc gaggagcttt
tcgatttcat 480agcttctgcg ctaggaaaat ttgcacaaaa ggaaggtggt aattttgagt
tgcaacaggg 540acggacaagg gaaataggat tcacgttttc ttttccggtg aaacagactt
caataagaac 600tggaatccta atcaaatgga caaaaggttt tgctgtctct ggaactgcgg
gaaaagatgt 660tgttgcttgt ctgaatgaag ccatggagag gcggggaatg gatatgcaag
tgtctgccct 720ggtcaatgac actgtgggaa cacttgccgg agcaagatac tgggatgatg
atgccatggt 780tgctgtcatt cttgggactg gaaccaatgc ttgctatgta gaacgtgtgg
atgctattcc 840taagctggca aaaaggatgt ctaagtctcc aataacgatt gtgaataccg
aatggggagc 900tttctcaaat ggccttcctt taactgagtt tgatagagaa atggatgccg
agagcattaa 960ccctggtgag cagatttttg aaaagaccat ctctggtatg taccttgggg
aaattgtgag 1020acgggtgctg gtcaaaatgg ccaaggttgg gggcttattt ggtagcagct
atgttccgga 1080gaagctagtc actccatttg tgctgaggac acctgatata tgtgccatgc
agcaggatac 1140atcaatagat cttgaagctg ttgagtctgt cctctatgat gtagctgggg
taaaatccga 1200tctaagtgca aggaaaacag tcgtggacat ttgcgatact attgcaaaac
gagggggtcg 1260tctagctggt gcaggaattg ttggtatatt acagaaaatg gaggaagatt
ctaaaggtct 1320catctttggt aaaagaacag ttgtagcaat ggatggaggc ttatatgagc
actatcctca 1380gtacagagga tacctccaag aagctgtcac agagctacta ggctcggaaa
tttcgaaaaa 1440tgtagtgata gaacattcaa aagatggatc tggtattgga gctgcattat
tagctgctgc 1500aaattcaaag tatgaacatg atgattaagg gaaaagctaa atttcatgtg
ttcaaaggtt 1560tgcttgagga aaaatacatg agcatttgta atttagaatg ttctaaagaa
ggattgatga 1620ttggatttat agaggctctt gtttgtgatg ccattttttg tactcccatt
aatgaagtac 1680gaagaagtga ggatttatat agccaacccc aatttatttg gaatcgaggc
gtagttgtaa 1740ttgtattata tgcagcactg acttatttat gtga
177430499PRTSolanum lycopersicum 30Met Ser Val Thr Val Ser Ser
Pro Ala Val Arg Ser Phe His Val Ser 1 5
10 15 Arg Ser Pro His Lys Thr Ile Ser Arg Pro Arg
Val Ile Ile Ser Ala 20 25
30 Val Arg Ser Thr Asp Ser Leu Gly Val Ala Pro Ile Leu Thr Lys
Leu 35 40 45 Gln
Lys Asp Cys Ala Thr Pro Leu Pro Val Leu Arg His Val Ala Asp 50
55 60 Ala Met Ala Asp Asp Met
Arg Ala Gly Leu Ala Val Asp Gly Gly Ser 65 70
75 80 Asp Leu Lys Met Ile Leu Ser Tyr Val Asp Thr
Leu Pro Thr Gly Asn 85 90
95 Glu Lys Gly Leu Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg
100 105 110 Val Leu
Arg Val Gln Leu Gly Gly Lys Glu Glu Arg Val Val Ala Thr 115
120 125 Glu Phe Glu Gln Val Ser Ile
Pro Gln Glu Leu Met Phe Ala Thr Ser 130 135
140 Glu Glu Leu Phe Asp Phe Ile Ala Ser Ala Leu Gly
Lys Phe Ala Gln 145 150 155
160 Lys Glu Gly Gly Asn Phe Glu Leu Gln Gln Gly Arg Thr Arg Glu Ile
165 170 175 Gly Phe Thr
Phe Ser Phe Pro Val Lys Gln Thr Ser Ile Arg Thr Gly 180
185 190 Ile Leu Ile Lys Trp Thr Lys Gly
Phe Ala Val Ser Gly Thr Ala Gly 195 200
205 Lys Asp Val Val Ala Cys Leu Asn Glu Ala Met Glu Arg
Arg Gly Met 210 215 220
Asp Met Gln Val Ser Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala 225
230 235 240 Gly Ala Arg Tyr
Trp Asp Asp Asp Ala Met Val Ala Val Ile Leu Gly 245
250 255 Thr Gly Thr Asn Ala Cys Tyr Val Glu
Arg Val Asp Ala Ile Pro Lys 260 265
270 Leu Ala Lys Arg Met Ser Lys Ser Pro Ile Thr Ile Val Asn
Thr Glu 275 280 285
Trp Gly Ala Phe Ser Asn Gly Leu Pro Leu Thr Glu Phe Asp Arg Glu 290
295 300 Met Asp Ala Glu Ser
Ile Asn Pro Gly Glu Gln Ile Phe Glu Lys Thr 305 310
315 320 Ile Ser Gly Met Tyr Leu Gly Glu Ile Val
Arg Arg Val Leu Val Lys 325 330
335 Met Ala Lys Val Gly Gly Leu Phe Gly Ser Ser Tyr Val Pro Glu
Lys 340 345 350 Leu
Val Thr Pro Phe Val Leu Arg Thr Pro Asp Ile Cys Ala Met Gln 355
360 365 Gln Asp Thr Ser Ile Asp
Leu Glu Ala Val Glu Ser Val Leu Tyr Asp 370 375
380 Val Ala Gly Val Lys Ser Asp Leu Ser Ala Arg
Lys Thr Val Val Asp 385 390 395
400 Ile Cys Asp Thr Ile Ala Lys Arg Gly Gly Arg Leu Ala Gly Ala Gly
405 410 415 Ile Val
Gly Ile Leu Gln Lys Met Glu Glu Asp Ser Lys Gly Leu Ile 420
425 430 Phe Gly Lys Arg Thr Val Val
Ala Met Asp Gly Gly Leu Tyr Glu His 435 440
445 Tyr Pro Gln Tyr Arg Gly Tyr Leu Gln Glu Ala Val
Thr Glu Leu Leu 450 455 460
Gly Ser Glu Ile Ser Lys Asn Val Val Ile Glu His Ser Lys Asp Gly 465
470 475 480 Ser Gly Ile
Gly Ala Ala Leu Leu Ala Ala Ala Asn Ser Lys Tyr Glu 485
490 495 His Asp Asp 311793DNASolanum
chacoense 31aatcccaatt ccccttctct atcatttttt ccttaatcgg atagtaagga
tgaagaaggc 60gacggtggct gcggtggtgg taggtacagc ggcggcggta gctgtggcgg
cgctcatcat 120gcgccaccgc atgggtaaat cgagcaaatg ggcacgtgcc agggcaattc
tgaaggaatt 180cgaggagaaa tgtgccaccc cagatggcaa gctgaagcaa gtggctgatg
ccatgacggt 240ggagatgcac gccggactcg cctctgaagg cggcagcaag ctcaagatgc
ttattagcta 300tgtcgataat ctccctactg gcgatgaagg aggagtcttt tatgcattgg
atcttggtgg 360aacaaatttt cgagtattgc gggtgcaatt ggggggaaaa gatggtggca
ttatccatca 420agaatttgcg gaggcatcaa ttcctccaaa tttgatggtt ggaacttcag
aagcactttt 480tgactatatt gcggcagaac ttgcaaaatt cgtagctgaa gaaggagaag
agtttcatcc 540acctcctggt aggcagagag aattaggctt caccttctcg ttcccaataa
tgcagacttc 600aatcaattct ggaactctta tcaggtggac gaaaggtttc tccattgatg
acacggttgg 660caaagatgtt gttgcagaac tgacaaaagc aatgcaaaaa cgagaaattg
atatgagggt 720ctcagcgctt gtgaatgata ctgttggaac attggctggt ggtagattca
ccaataagga 780tgtatccatt gctgtgatat taggtactgg gaccaatgca gcatatgtgg
aacgggctca 840ggcaattccc aaatggcacg gtcctctgcc taaatctgga gaaatggtga
tcaatatgga 900atggggtaac tttaggtcct cccaccttcc cttgacagag tacgatcatg
ctatggatac 960cgatagttta aatcctggtg aacagatatt tgagaagata tgttctggca
tgtacttggg 1020agaaatttta cgcagagttc tacttagaat ggctgaagaa gctggcattt
ttggcgagga 1080agttcctcca aaactcaaga attcattcat attgaggaca cctgaaatgt
ctgctatgca 1140tcatgacaca tcctctgatt tgagagtggt tggcgacaag ttgaaggata
tcttagagat 1200atccaatacc tccttgaaga caaggagatt agttgttgag ctgtgtaata
ttgttgcaac 1260acgtggcgcc agacttgcgg cagctgggat cttgggcatt atcaaaaaga
tgggaaagga 1320tacacccagg gaaagtggtc cagaaaagat tgtcgtagcc atggatggcg
gattgtatga 1380acattataca gaatacagta agtgcttgga gaacactttg gttgaattac
ttggaaagga 1440aatggccaca agtattgttt tcaagcacgc gaatgatggt tctggcattg
gcgctgcact 1500ccttgcggcc tctaactccg tgtatgttga agacaagtga gagtgatcaa
aatgatattt 1560agctagagga aactccattt accttttata ttatgttttt tctctggcct
tttctttttg 1620gattctctct ctcctttttg gttcttctta caattatttc cagaattttc
cagcacttgt 1680acttggtttt ctaggatatt attcctggga atgccaacat atttcttctg
gaaaagggtg 1740caaagaaatt atatgaaact cagcattgct tattcaaaaa aaaaaaaaaa
aaa 179332496PRTSolanum chacoense 32Met Lys Lys Ala Thr Val Ala
Ala Val Val Val Gly Thr Ala Ala Ala 1 5
10 15 Val Ala Val Ala Ala Leu Ile Met Arg His Arg
Met Gly Lys Ser Ser 20 25
30 Lys Trp Ala Arg Ala Arg Ala Ile Leu Lys Glu Phe Glu Glu Lys
Cys 35 40 45 Ala
Thr Pro Asp Gly Lys Leu Lys Gln Val Ala Asp Ala Met Thr Val 50
55 60 Glu Met His Ala Gly Leu
Ala Ser Glu Gly Gly Ser Lys Leu Lys Met 65 70
75 80 Leu Ile Ser Tyr Val Asp Asn Leu Pro Thr Gly
Asp Glu Gly Gly Val 85 90
95 Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Arg Val
100 105 110 Gln Leu
Gly Gly Lys Asp Gly Gly Ile Ile His Gln Glu Phe Ala Glu 115
120 125 Ala Ser Ile Pro Pro Asn Leu
Met Val Gly Thr Ser Glu Ala Leu Phe 130 135
140 Asp Tyr Ile Ala Ala Glu Leu Ala Lys Phe Val Ala
Glu Glu Gly Glu 145 150 155
160 Glu Phe His Pro Pro Pro Gly Arg Gln Arg Glu Leu Gly Phe Thr Phe
165 170 175 Ser Phe Pro
Ile Met Gln Thr Ser Ile Asn Ser Gly Thr Leu Ile Arg 180
185 190 Trp Thr Lys Gly Phe Ser Ile Asp
Asp Thr Val Gly Lys Asp Val Val 195 200
205 Ala Glu Leu Thr Lys Ala Met Gln Lys Arg Glu Ile Asp
Met Arg Val 210 215 220
Ser Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala Gly Gly Arg Phe 225
230 235 240 Thr Asn Lys Asp
Val Ser Ile Ala Val Ile Leu Gly Thr Gly Thr Asn 245
250 255 Ala Ala Tyr Val Glu Arg Ala Gln Ala
Ile Pro Lys Trp His Gly Pro 260 265
270 Leu Pro Lys Ser Gly Glu Met Val Ile Asn Met Glu Trp Gly
Asn Phe 275 280 285
Arg Ser Ser His Leu Pro Leu Thr Glu Tyr Asp His Ala Met Asp Thr 290
295 300 Asp Ser Leu Asn Pro
Gly Glu Gln Ile Phe Glu Lys Ile Cys Ser Gly 305 310
315 320 Met Tyr Leu Gly Glu Ile Leu Arg Arg Val
Leu Leu Arg Met Ala Glu 325 330
335 Glu Ala Gly Ile Phe Gly Glu Glu Val Pro Pro Lys Leu Lys Asn
Ser 340 345 350 Phe
Ile Leu Arg Thr Pro Glu Met Ser Ala Met His His Asp Thr Ser 355
360 365 Ser Asp Leu Arg Val Val
Gly Asp Lys Leu Lys Asp Ile Leu Glu Ile 370 375
380 Ser Asn Thr Ser Leu Lys Thr Arg Arg Leu Val
Val Glu Leu Cys Asn 385 390 395
400 Ile Val Ala Thr Arg Gly Ala Arg Leu Ala Ala Ala Gly Ile Leu Gly
405 410 415 Ile Ile
Lys Lys Met Gly Lys Asp Thr Pro Arg Glu Ser Gly Pro Glu 420
425 430 Lys Ile Val Val Ala Met Asp
Gly Gly Leu Tyr Glu His Tyr Thr Glu 435 440
445 Tyr Ser Lys Cys Leu Glu Asn Thr Leu Val Glu Leu
Leu Gly Lys Glu 450 455 460
Met Ala Thr Ser Ile Val Phe Lys His Ala Asn Asp Gly Ser Gly Ile 465
470 475 480 Gly Ala Ala
Leu Leu Ala Ala Ser Asn Ser Val Tyr Val Glu Asp Lys 485
490 495 331619DNASolanum tuberosum
33taaattccgg caaaaaaaat gaagaaagtg acggtgggag ccgccgtggt tggtgcagct
60gcggtgtgtg cagtggcggc gttaatagtg aaccaccgga tgcggaaatc tagtaaatgg
120ggtcgtgcta tggctattct tcgtgaattt gaggaaaagt gtaagactca agatgcaaag
180cttaagcaag ttgctgatgc tatgactgtt gagatgcacg ccggacttgc ttctgaaggc
240ggccagagct caagatgctt atcacctatg tcgataatct cccaactggt gatgaagctg
300ggcgtctttt atgcattgga tcttggtgga acaaattttc gagtattgcg agtgcaattg
360ggtggaaaag atggtggtat tattcatcaa gaatttgctg aggcatcaat tcctccaagt
420ttgatggttg ggacttcaga tgcacttttt gattatattg cggctgagct tgctaaattt
480gttgctgcgg aagaggaaaa atttcatcaa cctcctggta agcagagaga actaggtttt
540caccttctca ttcccagtaa tgcagacttc aataactctg ggactattat gcggtggaca
600aaaggcttct caattgatga tgcggttggc caagatgttg ttggagaact cacaaaagct
660atgaaagaaa aggtgctcga tatgcgggtc tcagctctgg tgaatgatac tgttggaaca
720ttagctggtg gtaaatatac gcaaaaggat gtagctgttg ctgttatctt aggtacaggg
780acgaatgcag cttatgtgga gcgggtgcag gcaattccaa agtggcatgg tcctgtgcca
840aaatctggtg aaatggttat caatatggaa tggggtaatt ttaggtcatc ccatctgccg
900ttgacagagt atgatcatgc gttggataac gagagtttaa atcctgctga acagatattt
960gagaagatga cttctggcat gtacttggga gaaattttac gcagagttct cactagggtg
1020gctgaagaag tcctggcgtt tttggcgatg aggtccctcc aaagcctcaa ggattcattt
1080gtgttaagga cacctgatat gtctgctatg catcatgaca catcccctga tctgaaagtg
1140gttggtgaaa agctgaagga tattttagag atatctaata cctccttgaa gacaaggaaa
1200ttagtgttga gcctgtgcaa tatcgttgca acacgtgggg caagacttga cgctgcaggg
1260gtattgggca tcttgaaaaa gatgggaaga gatacgccta agcagggtgg ttcagaaagg
1320acggttatag ccatggatgg cgggttgtat gagcactata cagaatatag aatgtgttta
1380gagaactctt tgaaggactt gctcggagag gaattggcaa cgagcatcgt ttttgtgcac
1440tccaatgatg gttctggcat tggtgctgct cttcttcgtg cctctcattc aatgtacctt
1500gaagatcaag cttaggcgag tgtgatcaaa tccataactt gctagagggg actaccttta
1560gactctatat tttgctctca agcgttttcg gtattttctc tcctttattg taccatcaa
161934498PRTSolanum tuberosum 34Met Lys Lys Val Thr Val Gly Ala Ala Val
Val Gly Ala Ala Ala Val 1 5 10
15 Cys Ala Val Ala Ala Leu Ile Val Asn His Arg Met Arg Lys Ser
Ser 20 25 30 Lys
Trp Gly Arg Ala Met Ala Ile Leu Arg Glu Phe Glu Glu Lys Cys 35
40 45 Lys Thr Gln Asp Ala Lys
Leu Lys Gln Val Ala Asp Ala Met Thr Val 50 55
60 Glu Met His Ala Gly Leu Ala Ser Glu Gly Gly
Gln Ser Ser Arg Cys 65 70 75
80 Leu Ser Pro Met Ser Ile Ile Ser Gln Leu Val Met Lys Leu Gly Val
85 90 95 Phe Tyr
Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Arg Val 100
105 110 Gln Leu Gly Gly Lys Asp Gly
Gly Ile Ile His Gln Glu Phe Ala Glu 115 120
125 Ala Ser Ile Pro Pro Ser Leu Met Val Gly Thr Ser
Asp Ala Leu Phe 130 135 140
Asp Tyr Ile Ala Ala Glu Leu Ala Lys Phe Val Ala Ala Glu Glu Glu 145
150 155 160 Lys Phe His
Gln Pro Pro Gly Lys Gln Arg Glu Leu Gly Phe His Leu 165
170 175 Leu Ile Pro Ser Asn Ala Asp Phe
Asn Asn Ser Gly Thr Ile Met Arg 180 185
190 Trp Thr Lys Gly Phe Ser Ile Asp Asp Ala Val Gly Gln
Asp Val Val 195 200 205
Gly Glu Leu Thr Lys Ala Met Lys Glu Lys Val Leu Asp Met Arg Val 210
215 220 Ser Ala Leu Val
Asn Asp Thr Val Gly Thr Leu Ala Gly Gly Lys Tyr 225 230
235 240 Thr Gln Lys Asp Val Ala Val Ala Val
Ile Leu Gly Thr Gly Thr Asn 245 250
255 Ala Ala Tyr Val Glu Arg Val Gln Ala Ile Pro Lys Trp His
Gly Pro 260 265 270
Val Pro Lys Ser Gly Glu Met Val Ile Asn Met Glu Trp Gly Asn Phe
275 280 285 Arg Ser Ser His
Leu Pro Leu Thr Glu Tyr Asp His Ala Leu Asp Asn 290
295 300 Glu Ser Leu Asn Pro Ala Glu Gln
Ile Phe Glu Lys Met Thr Ser Gly 305 310
315 320 Met Tyr Leu Gly Glu Ile Leu Arg Arg Val Leu Thr
Arg Val Ala Glu 325 330
335 Glu Val Leu Ala Phe Leu Ala Met Arg Ser Leu Gln Ser Leu Lys Asp
340 345 350 Ser Phe Val
Leu Arg Thr Pro Asp Met Ser Ala Met His His Asp Thr 355
360 365 Ser Pro Asp Leu Lys Val Val Gly
Glu Lys Leu Lys Asp Ile Leu Glu 370 375
380 Ile Ser Asn Thr Ser Leu Lys Thr Arg Lys Leu Val Leu
Ser Leu Cys 385 390 395
400 Asn Ile Val Ala Thr Arg Gly Ala Arg Leu Asp Ala Ala Gly Val Leu
405 410 415 Gly Ile Leu Lys
Lys Met Gly Arg Asp Thr Pro Lys Gln Gly Gly Ser 420
425 430 Glu Arg Thr Val Ile Ala Met Asp Gly
Gly Leu Tyr Glu His Tyr Thr 435 440
445 Glu Tyr Arg Met Cys Leu Glu Asn Ser Leu Lys Asp Leu Leu
Gly Glu 450 455 460
Glu Leu Ala Thr Ser Ile Val Phe Val His Ser Asn Asp Gly Ser Gly 465
470 475 480 Ile Gly Ala Ala Leu
Leu Arg Ala Ser His Ser Met Tyr Leu Glu Asp 485
490 495 Gln Ala 351874DNASolanum tuberosum
35ttcggcacga gatcaatccc aattcccctt ctctatcatt ttttccttaa tcggatagta
60aggatgaaga aggcgacggt gggtgcggtg gtcgtaggta cagcggcggc ggtagctgtg
120gcggcgctca tcatgcgcca ccgcatgggt aaatcgagca aatgggcacg tgccagggca
180attctgaagg aattcgagga gaaatgtgcc accccagatg gcaagctgaa gcaagtggct
240gatgccatga cggtggagat gcacgccgga ctcgcctctg aaggcggcag caagctcaag
300atgcttatta gctatgtcga taatctccct actggcgatg aaggaggagt cttttatgca
360ttggatcttg gtggaacaaa ttttcgagta ttgcgggtgc aattgggggg aaaagatggt
420ggcattatcc atcaagaatt tgcggaggca tcaattcctc caaatttgat ggttggaact
480tcagaagcac tttttgacta tattgcggca gaacttgcaa aattcgtagc tgaagaagga
540gaagagtttc atccacctcc tggtaggcag agagaattag gcttcacctt ctcgttccca
600ataatgcaaa cttcaatcaa ttctggaact cttatcaggt ggacgaaagg tttctccatt
660gatgacacgg ttggcaaaga tgttgttgca gaactgacaa aagcaatgca aaaacgagaa
720attgatatga gggtctcagc gcttgtgaat gatactgttg gaacattggc tggtggtaga
780ttcaccaata aggatgtatc cattgctgtg atattaggta ctgggaccaa tgcagcatat
840gtggaacggg ctcaggcaat tcccaaatgg cacggtcctc tgcctaaatc tggagaaatg
900gtgatcaata tggaatgggg taactttagg tcctcccacc ttcccttgac agagtacgat
960catgctatgg ataccaatag tttaaatcct ggtgaacaga tatttgagaa gatatgttct
1020ggcatgtact tgggagaaat tttacgcaga gttctactta gaatggctga agaagctggc
1080atttttggcg aggaagttcc tccaaaactc aagaattcat tcatattgag gacacctgaa
1140atgtctgcta tgcatcatga cacatcctct gatttgagag tggttggcga caagttgaag
1200gatatcttag agatctccaa ttcctccttg aagacaagga gattagttgt tgagctgtgt
1260aatattgttg caacacgtgg cgccagactt gcagcagctg ggatcttggg cattatcaaa
1320aagatgggaa aggatacacc cagggaaagt ggtccagaaa agattgtcgt agccatggat
1380ggcggattgt atgaacatta tacagaatac agtaagtgct tggagaacac tttggttgaa
1440ttgcttggaa aggaaatggc cacaagtatt gttttcaagc acgcgaatga tggttctggc
1500attggcgcgg cactccttgc ggcctctaac tccgtgtatg ttgaagacaa gtgagagtga
1560tcaaaatgat atttagctag aggaaactcc atttaccttc tatattatgt tttttctcta
1620gccttttctt tttggattct ctctcctttt tggttcttct tacaattatt tccagaattt
1680tccagtactt gtacttggtt ttctaggata ttattcctgg gaatgccaac atatttcttc
1740tggaaaaggg tgcaaagaaa atatatatga aacgcagcac tgcttattct cccctttggt
1800ctttagtttc caactgcaat taaattttgt agggttgaat aaaatggtgt acttttcaaa
1860aaaaaaaaaa aaaa
187436496PRTSolanum tuberosum 36Met Lys Lys Ala Thr Val Gly Ala Val Val
Val Gly Thr Ala Ala Ala 1 5 10
15 Val Ala Val Ala Ala Leu Ile Met Arg His Arg Met Gly Lys Ser
Ser 20 25 30 Lys
Trp Ala Arg Ala Arg Ala Ile Leu Lys Glu Phe Glu Glu Lys Cys 35
40 45 Ala Thr Pro Asp Gly Lys
Leu Lys Gln Val Ala Asp Ala Met Thr Val 50 55
60 Glu Met His Ala Gly Leu Ala Ser Glu Gly Gly
Ser Lys Leu Lys Met 65 70 75
80 Leu Ile Ser Tyr Val Asp Asn Leu Pro Thr Gly Asp Glu Gly Gly Val
85 90 95 Phe Tyr
Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Arg Val 100
105 110 Gln Leu Gly Gly Lys Asp Gly
Gly Ile Ile His Gln Glu Phe Ala Glu 115 120
125 Ala Ser Ile Pro Pro Asn Leu Met Val Gly Thr Ser
Glu Ala Leu Phe 130 135 140
Asp Tyr Ile Ala Ala Glu Leu Ala Lys Phe Val Ala Glu Glu Gly Glu 145
150 155 160 Glu Phe His
Pro Pro Pro Gly Arg Gln Arg Glu Leu Gly Phe Thr Phe 165
170 175 Ser Phe Pro Ile Met Gln Thr Ser
Ile Asn Ser Gly Thr Leu Ile Arg 180 185
190 Trp Thr Lys Gly Phe Ser Ile Asp Asp Thr Val Gly Lys
Asp Val Val 195 200 205
Ala Glu Leu Thr Lys Ala Met Gln Lys Arg Glu Ile Asp Met Arg Val 210
215 220 Ser Ala Leu Val
Asn Asp Thr Val Gly Thr Leu Ala Gly Gly Arg Phe 225 230
235 240 Thr Asn Lys Asp Val Ser Ile Ala Val
Ile Leu Gly Thr Gly Thr Asn 245 250
255 Ala Ala Tyr Val Glu Arg Ala Gln Ala Ile Pro Lys Trp His
Gly Pro 260 265 270
Leu Pro Lys Ser Gly Glu Met Val Ile Asn Met Glu Trp Gly Asn Phe
275 280 285 Arg Ser Ser His
Leu Pro Leu Thr Glu Tyr Asp His Ala Met Asp Thr 290
295 300 Asn Ser Leu Asn Pro Gly Glu Gln
Ile Phe Glu Lys Ile Cys Ser Gly 305 310
315 320 Met Tyr Leu Gly Glu Ile Leu Arg Arg Val Leu Leu
Arg Met Ala Glu 325 330
335 Glu Ala Gly Ile Phe Gly Glu Glu Val Pro Pro Lys Leu Lys Asn Ser
340 345 350 Phe Ile Leu
Arg Thr Pro Glu Met Ser Ala Met His His Asp Thr Ser 355
360 365 Ser Asp Leu Arg Val Val Gly Asp
Lys Leu Lys Asp Ile Leu Glu Ile 370 375
380 Ser Asn Ser Ser Leu Lys Thr Arg Arg Leu Val Val Glu
Leu Cys Asn 385 390 395
400 Ile Val Ala Thr Arg Gly Ala Arg Leu Ala Ala Ala Gly Ile Leu Gly
405 410 415 Ile Ile Lys Lys
Met Gly Lys Asp Thr Pro Arg Glu Ser Gly Pro Glu 420
425 430 Lys Ile Val Val Ala Met Asp Gly Gly
Leu Tyr Glu His Tyr Thr Glu 435 440
445 Tyr Ser Lys Cys Leu Glu Asn Thr Leu Val Glu Leu Leu Gly
Lys Glu 450 455 460
Met Ala Thr Ser Ile Val Phe Lys His Ala Asn Asp Gly Ser Gly Ile 465
470 475 480 Gly Ala Ala Leu Leu
Ala Ala Ser Asn Ser Val Tyr Val Glu Asp Lys 485
490 495 372300DNANicotiana tabacum 37gtgaattgct
acattattac ggcaactatg aatatgaaag taaggaacag tcggtagtta 60gctaaccata
ccttccaacg caacaataac ggcgtacaac agtagtatag taatattcat 120cgtttaaatc
cttacatata tttcatcatt aatttattca aatctaactg ttcgcttacc 180atcattacga
tataaaaaag aaaatttata tagtgttaaa gattcacaac tgtataaact 240ctctgtttca
gcatcaccat ttattgtctc ttcttttttt ctccgtcaca cgttcctttt 300gatctcactg
ttaattatgt cggtcaccgt tagctcgccg gccggccgat ccttccatat 360ttcacgatca
ccttacaaaa agatctccaa gccacgtgtc attatcgctg ccgtccgatc 420tggtgttagt
ttagcggtag caccaatatt gactaagttg cagaaagact gtgcaactcc 480acttcctgtt
ttgcgccacg tggctgatgc catggccgtt gatatgcggg ctggacttgc 540cgtcgatggt
ggcagtgatc tgaagatgat ccttagttat attgacactt taccaactgg 600gaatgagaaa
ggtttgttct atgcattgga ccttggtggt acaaatttcc gagtgttaag 660agtgcagtta
ggtggtaaag aagagcgcgt aatcgccact gagtttgagc aagtctctat 720acctcaagaa
ttgatgtttg caacctctga ggagttgttc gatttcatag cttctgagct 780aggaaaattt
tcacaaagtg aaggcggtaa gtttgagatg caacaaggaa ggactagaga 840aataggattc
acattttctt tcccagtgaa gcagacttca gttaaatctg gcatcctaat 900caagtggaca
aagggttttg cagtctctgg aactgcagga aaagatgtgg ttgcttgttt 960aaatgaagcc
atggaaaggc agggattggg aatgcaagtc tcggccctgg tcaatgacac 1020tgtagcaaca
cttgctggag cgagatactg ggacaatgat gtcatggttg ctgtcattct 1080tgggactgga
accaatgctt gctacgtaga acgtgtggat gctattccta aactgccaca 1140aaggatgtcc
aactctccag aaacaattgt gaatactgaa tggggagcat tttcaaatgg 1200ccttccttta
actgagtttg atagagaaat ggatgccgag agcattaacc ctggtgagca 1260gatttttgag
aaaacaatct ctggtatgta ccttggagaa attgtaagac gggtgctggt 1320caaaatggcc
aaggttggcg gcttatttgg cggtggctat gttccagaaa agttagtcac 1380tccatttgtg
ctgaggacac cggatatatg tgcaatgcag caggatacat ccagagatct 1440tgaagctgtt
gagtctgtcc tctatgatat agctggggta aaatctgatc taagtgcaag 1500gaaaacagtc
gtagacattt gcgatactat tgcaaatcga ggggggcgtc tagctggtgc 1560aggaattgtt
gggattctcc agaaaatgga agaggattca aaaggcgtaa tcttcggtaa 1620gagaacagtt
gtagcaatgg atggaggttt atatgagcac tatcctcagt acagagaata 1680cctccaagaa
gctgtcacag aacttcttgg atcagaaatt tctaaaaatg tagtgataga 1740gcattcaaaa
gatggatctg gaattggagc tgcattatta gctgctgcaa actcaaagta 1800tgaacatgat
tattaggaaa atggtaattt tcatgtctaa aaatgctggt agagcatttg 1860taattttgtt
tttgcaattt agaatgtatt aagaaggatt gatcattgga tttctcattc 1920tttaagggct
cgtttggtac gagggataag agataattac atccgggatt aaatttgaga 1980taagtttatc
ccacgtttgg ttgggataaa atcgcggtat aactaatccc gggattagtt 2040attccgggat
tgtagtgtta tttttatccc tatgagatgg tgggataact aatcctaaga 2100taattaattc
tgggataatc tgtttcccaa ccaaacgatc cttaagaagt ggcttgttgt 2160attttatgcc
atttttgtat taccataatg cagcagtgaa gtatttatgt gctaaaagca 2220gatgtatcag
taaatataga atatcttgtt tgcttataaa ttgttaaata agcatatttc 2280tgtggtattt
gaggcggccg
230038499PRTNicotiana tabacum 38Met Ser Val Thr Val Ser Ser Pro Ala Gly
Arg Ser Phe His Ile Ser 1 5 10
15 Arg Ser Pro Tyr Lys Lys Ile Ser Lys Pro Arg Val Ile Ile Ala
Ala 20 25 30 Val
Arg Ser Gly Val Ser Leu Ala Val Ala Pro Ile Leu Thr Lys Leu 35
40 45 Gln Lys Asp Cys Ala Thr
Pro Leu Pro Val Leu Arg His Val Ala Asp 50 55
60 Ala Met Ala Val Asp Met Arg Ala Gly Leu Ala
Val Asp Gly Gly Ser 65 70 75
80 Asp Leu Lys Met Ile Leu Ser Tyr Ile Asp Thr Leu Pro Thr Gly Asn
85 90 95 Glu Lys
Gly Leu Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg 100
105 110 Val Leu Arg Val Gln Leu Gly
Gly Lys Glu Glu Arg Val Ile Ala Thr 115 120
125 Glu Phe Glu Gln Val Ser Ile Pro Gln Glu Leu Met
Phe Ala Thr Ser 130 135 140
Glu Glu Leu Phe Asp Phe Ile Ala Ser Glu Leu Gly Lys Phe Ser Gln 145
150 155 160 Ser Glu Gly
Gly Lys Phe Glu Met Gln Gln Gly Arg Thr Arg Glu Ile 165
170 175 Gly Phe Thr Phe Ser Phe Pro Val
Lys Gln Thr Ser Val Lys Ser Gly 180 185
190 Ile Leu Ile Lys Trp Thr Lys Gly Phe Ala Val Ser Gly
Thr Ala Gly 195 200 205
Lys Asp Val Val Ala Cys Leu Asn Glu Ala Met Glu Arg Gln Gly Leu 210
215 220 Gly Met Gln Val
Ser Ala Leu Val Asn Asp Thr Val Ala Thr Leu Ala 225 230
235 240 Gly Ala Arg Tyr Trp Asp Asn Asp Val
Met Val Ala Val Ile Leu Gly 245 250
255 Thr Gly Thr Asn Ala Cys Tyr Val Glu Arg Val Asp Ala Ile
Pro Lys 260 265 270
Leu Pro Gln Arg Met Ser Asn Ser Pro Glu Thr Ile Val Asn Thr Glu
275 280 285 Trp Gly Ala Phe
Ser Asn Gly Leu Pro Leu Thr Glu Phe Asp Arg Glu 290
295 300 Met Asp Ala Glu Ser Ile Asn Pro
Gly Glu Gln Ile Phe Glu Lys Thr 305 310
315 320 Ile Ser Gly Met Tyr Leu Gly Glu Ile Val Arg Arg
Val Leu Val Lys 325 330
335 Met Ala Lys Val Gly Gly Leu Phe Gly Gly Gly Tyr Val Pro Glu Lys
340 345 350 Leu Val Thr
Pro Phe Val Leu Arg Thr Pro Asp Ile Cys Ala Met Gln 355
360 365 Gln Asp Thr Ser Arg Asp Leu Glu
Ala Val Glu Ser Val Leu Tyr Asp 370 375
380 Ile Ala Gly Val Lys Ser Asp Leu Ser Ala Arg Lys Thr
Val Val Asp 385 390 395
400 Ile Cys Asp Thr Ile Ala Asn Arg Gly Gly Arg Leu Ala Gly Ala Gly
405 410 415 Ile Val Gly Ile
Leu Gln Lys Met Glu Glu Asp Ser Lys Gly Val Ile 420
425 430 Phe Gly Lys Arg Thr Val Val Ala Met
Asp Gly Gly Leu Tyr Glu His 435 440
445 Tyr Pro Gln Tyr Arg Glu Tyr Leu Gln Glu Ala Val Thr Glu
Leu Leu 450 455 460
Gly Ser Glu Ile Ser Lys Asn Val Val Ile Glu His Ser Lys Asp Gly 465
470 475 480 Ser Gly Ile Gly Ala
Ala Leu Leu Ala Ala Ala Asn Ser Lys Tyr Glu 485
490 495 His Asp Tyr 391494DNANicotiana tabacum
39atgaagaaag cgacggtggg agccgccgtg gttggcgccg ctacggtatg tgctgtggcg
60gcattgatag tgaaccaccg tatgcgcaaa tctagtaaat gggcacgtgc tatggctatt
120cttcgtgaat ttgaggaaaa gtgtgggacc cctgatgcta agctcaagca agtcgctgat
180gctatgaccg tcgagatgca cgctggactt gcctccgaag gtggtagcaa gctcaagatg
240cttatcactt acgtcgataa tctccccacc ggtgatgaag ccggcgtctt ttatgcgttg
300gatcttggtg gaacaaattt tcgagtattg cgggtgcagc ttggtggaaa agatggtggt
360attattcatc aagaatttgc agaggcatca attcctccaa atttgatggt tgggacttca
420gaagaacttt ttgattacat tgcggcagaa cttgcaaaat ttgtcgctga ggaagaggag
480aaatttcaac aacctcctgg taagcagaga gaactaggtt tcaccttctc attcccggta
540atgcagactt caatcaactc tgggactatt atgaggtgga caaagggctt ctccatcgat
600gatgcggttg gccaagatgt tgtcggagaa ctcacaaaag ctatgaaaag aaagggcgtt
660gatatgcggg tctcagctct ggtgaatgat actgttggga cgttggctgg tggtaaatat
720acacacaacg acgtagctgt tgctgttatc ttaggtacag ggaccaatgc agcttatgtg
780gaacgggtgc aggcgattcc aaagtggcat ggtcctatgc caaaatctgg tgaaatggtt
840atcaacatgg aatggggtaa ttttaggtca tcccatcttc ccttgacaca gtatgatcat
900gcgttggata ctaatagttt gaatcctggt gatcagatat ttgagaagat gacttctggc
960atgtacttgg gagaaatttt acgcagagtt ctactcagga tggccgaaga agctggcatt
1020tttggtgatg aggtccctcc aaagctcaag agtccatttg tattgaggac acctgatatg
1080tctgctatgc atcatgacac atcatctgat ctgagagtgg ttggtgacaa gctgaaggat
1140attttagaga tatctaatac ctcattgaag acaaggagat tagtcgttga gctgtgcaac
1200atcgttgcaa cacgaggggc aaggcttgca gctgcaggtg tattggggat cttgaagaag
1260atgggaaggg atacgcctag gcaaggtggt ccgcagaaga tggttgtagc catggatggc
1320ggattgtatg agcactatgc agagtacagg acgtgcttag agaacacatt gaaggaattg
1380cttggagatg aattggcgac aagcattgtt ttcgagcact ccaatgatgg ttctggcatt
1440ggtgctgctc ttcttgctgc ctctaactca atgtaccttg aagataagtc ctag
149440497PRTNicotiana tabacum 40Met Lys Lys Ala Thr Val Gly Ala Ala Val
Val Gly Ala Ala Thr Val 1 5 10
15 Cys Ala Val Ala Ala Leu Ile Val Asn His Arg Met Arg Lys Ser
Ser 20 25 30 Lys
Trp Ala Arg Ala Met Ala Ile Leu Arg Glu Phe Glu Glu Lys Cys 35
40 45 Gly Thr Pro Asp Ala Lys
Leu Lys Gln Val Ala Asp Ala Met Thr Val 50 55
60 Glu Met His Ala Gly Leu Ala Ser Glu Gly Gly
Ser Lys Leu Lys Met 65 70 75
80 Leu Ile Thr Tyr Val Asp Asn Leu Pro Thr Gly Asp Glu Ala Gly Val
85 90 95 Phe Tyr
Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Arg Val 100
105 110 Gln Leu Gly Gly Lys Asp Gly
Gly Ile Ile His Gln Glu Phe Ala Glu 115 120
125 Ala Ser Ile Pro Pro Asn Leu Met Val Gly Thr Ser
Glu Glu Leu Phe 130 135 140
Asp Tyr Ile Ala Ala Glu Leu Ala Lys Phe Val Ala Glu Glu Glu Glu 145
150 155 160 Lys Phe Gln
Gln Pro Pro Gly Lys Gln Arg Glu Leu Gly Phe Thr Phe 165
170 175 Ser Phe Pro Val Met Gln Thr Ser
Ile Asn Ser Gly Thr Ile Met Arg 180 185
190 Trp Thr Lys Gly Phe Ser Ile Asp Asp Ala Val Gly Gln
Asp Val Val 195 200 205
Gly Glu Leu Thr Lys Ala Met Lys Arg Lys Gly Val Asp Met Arg Val 210
215 220 Ser Ala Leu Val
Asn Asp Thr Val Gly Thr Leu Ala Gly Gly Lys Tyr 225 230
235 240 Thr His Asn Asp Val Ala Val Ala Val
Ile Leu Gly Thr Gly Thr Asn 245 250
255 Ala Ala Tyr Val Glu Arg Val Gln Ala Ile Pro Lys Trp His
Gly Pro 260 265 270
Met Pro Lys Ser Gly Glu Met Val Ile Asn Met Glu Trp Gly Asn Phe
275 280 285 Arg Ser Ser His
Leu Pro Leu Thr Gln Tyr Asp His Ala Leu Asp Thr 290
295 300 Asn Ser Leu Asn Pro Gly Asp Gln
Ile Phe Glu Lys Met Thr Ser Gly 305 310
315 320 Met Tyr Leu Gly Glu Ile Leu Arg Arg Val Leu Leu
Arg Met Ala Glu 325 330
335 Glu Ala Gly Ile Phe Gly Asp Glu Val Pro Pro Lys Leu Lys Ser Pro
340 345 350 Phe Val Leu
Arg Thr Pro Asp Met Ser Ala Met His His Asp Thr Ser 355
360 365 Ser Asp Leu Arg Val Val Gly Asp
Lys Leu Lys Asp Ile Leu Glu Ile 370 375
380 Ser Asn Thr Ser Leu Lys Thr Arg Arg Leu Val Val Glu
Leu Cys Asn 385 390 395
400 Ile Val Ala Thr Arg Gly Ala Arg Leu Ala Ala Ala Gly Val Leu Gly
405 410 415 Ile Leu Lys Lys
Met Gly Arg Asp Thr Pro Arg Gln Gly Gly Pro Gln 420
425 430 Lys Met Val Val Ala Met Asp Gly Gly
Leu Tyr Glu His Tyr Ala Glu 435 440
445 Tyr Arg Thr Cys Leu Glu Asn Thr Leu Lys Glu Leu Leu Gly
Asp Glu 450 455 460
Leu Ala Thr Ser Ile Val Phe Glu His Ser Asn Asp Gly Ser Gly Ile 465
470 475 480 Gly Ala Ala Leu Leu
Ala Ala Ser Asn Ser Met Tyr Leu Glu Asp Lys 485
490 495 Ser 411497DNAHelianthus annuus
41atgggtaagg tagcagtagc ggccacggtg gtctgcgccg ccgctgtgac cgcggcggcg
60gtggttgtgg tccggcaccg gatgaagaac tccggcaagt gggcaaaagc tatggagatt
120ttgaaggagt ttgaggacaa gtgtggaact ccggtttcaa aactccggca ggttgctgac
180gccatgacgg tggagatgca cgccggactt gcttccgacg gtggtagtaa actcaagatg
240ttaatcagct atgttgacaa tcttcccact ggggacgaaa cgggaatttt ctatgccctt
300gatcttggtg gtacaaactt tcgtgttctt cgtgtgaaat taggcggagt aggaaacgtg
360aaaaaagaat ttaaagaagt ttcaatccct ccgaatctca tgatcgggaa atctgaggat
420ttatttgatt ttattgcggg agaacttgca aaattcgtgg ctactgagga tgaagatatg
480cagattccac ctggcacgca gcgagaatta gggtttacct tttcgtttcc ggttaaacaa
540tcatcaattg cgggagggac tcttgtaaga tggacaaaag gcttcaatat cgaagatgca
600gttggggcgg atgttgtaga agagttgaca aaagcgatgg aaagggctgg ccttgatatg
660cgtgtttcgg ctttggtgaa tgatacagtt ggaacgttag ctggagggcg atatggtaat
720tccgatgtca ttgctgctgt aatattaggt acaggaacta atgcagcata tgtggagcga
780gcgaatgcaa tccctaaatg gcaaggtctt cttcctaaat caggagagat ggttataaac
840atggaatggg gcaacttccg gtcatcacat cttcctttga ccgagtatga tgaaggtctt
900gatggtgata gcctgaaccc tggagagcag atatatgaga aactgatttc cggaatgtat
960ttgggggaag ttgtgagacg agtcttgtta aagatggcgg aagaagccga gttttttgga
1020gatattgtgc catccaaact ccaaaagccc tttatattaa ggacccctga tatgtctgca
1080atgcatcatg attctactcc ggatctcaaa gtggttgcaa ccaagttgaa agatatcctc
1140gagatatcca acacttctct aaagatgaga aaagtaattg tggaagtctg tgaccttgtg
1200gcaactcgtg gtgctcgtct ctcagcggct ggaatcctag gaatcctcaa aaaaattgga
1260aaagatactt gcaaggaagg agaagaaaac cagaaatcag taatagcaat ggacggcggg
1320ctgtttgaac actacaccaa gttccgaaaa acaatgcaag ataccatgaa tgaattgtta
1380ggtgaagaaa tttccaaaaa catcattgtc gagctatcga atgacgggtc gggtctcggt
1440gcggctcttc ttgcagcgtc tcactcacag tatctcgaat atatagagtc cacataa
149742498PRTHelianthus annuus 42Met Gly Lys Val Ala Val Ala Ala Thr Val
Val Cys Ala Ala Ala Val 1 5 10
15 Thr Ala Ala Ala Val Val Val Val Arg His Arg Met Lys Asn Ser
Gly 20 25 30 Lys
Trp Ala Lys Ala Met Glu Ile Leu Lys Glu Phe Glu Asp Lys Cys 35
40 45 Gly Thr Pro Val Ser Lys
Leu Arg Gln Val Ala Asp Ala Met Thr Val 50 55
60 Glu Met His Ala Gly Leu Ala Ser Asp Gly Gly
Ser Lys Leu Lys Met 65 70 75
80 Leu Ile Ser Tyr Val Asp Asn Leu Pro Thr Gly Asp Glu Thr Gly Ile
85 90 95 Phe Tyr
Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Arg Val 100
105 110 Lys Leu Gly Gly Val Gly Asn
Val Lys Lys Glu Phe Lys Glu Val Ser 115 120
125 Ile Pro Pro Asn Leu Met Ile Gly Lys Ser Glu Asp
Leu Phe Asp Phe 130 135 140
Ile Ala Gly Glu Leu Ala Lys Phe Val Ala Thr Glu Asp Glu Asp Met 145
150 155 160 Gln Ile Pro
Pro Gly Thr Gln Arg Glu Leu Gly Phe Thr Phe Ser Phe 165
170 175 Pro Val Lys Gln Ser Ser Ile Ala
Gly Gly Thr Leu Val Arg Trp Thr 180 185
190 Lys Gly Phe Asn Ile Glu Asp Ala Val Gly Ala Asp Val
Val Glu Glu 195 200 205
Leu Thr Lys Ala Met Glu Arg Ala Gly Leu Asp Met Arg Val Ser Ala 210
215 220 Leu Val Asn Asp
Thr Val Gly Thr Leu Ala Gly Gly Arg Tyr Gly Asn 225 230
235 240 Ser Asp Val Ile Ala Ala Val Ile Leu
Gly Thr Gly Thr Asn Ala Ala 245 250
255 Tyr Val Glu Arg Ala Asn Ala Ile Pro Lys Trp Gln Gly Leu
Leu Pro 260 265 270
Lys Ser Gly Glu Met Val Ile Asn Met Glu Trp Gly Asn Phe Arg Ser
275 280 285 Ser His Leu Pro
Leu Thr Glu Tyr Asp Glu Gly Leu Asp Gly Asp Ser 290
295 300 Leu Asn Pro Gly Glu Gln Ile Tyr
Glu Lys Leu Ile Ser Gly Met Tyr 305 310
315 320 Leu Gly Glu Val Val Arg Arg Val Leu Leu Lys Met
Ala Glu Glu Ala 325 330
335 Glu Phe Phe Gly Asp Ile Val Pro Ser Lys Leu Gln Lys Pro Phe Ile
340 345 350 Leu Arg Thr
Pro Asp Met Ser Ala Met His His Asp Ser Thr Pro Asp 355
360 365 Leu Lys Val Val Ala Thr Lys Leu
Lys Asp Ile Leu Glu Ile Ser Asn 370 375
380 Thr Ser Leu Lys Met Arg Lys Val Ile Val Glu Val Cys
Asp Leu Val 385 390 395
400 Ala Thr Arg Gly Ala Arg Leu Ser Ala Ala Gly Ile Leu Gly Ile Leu
405 410 415 Lys Lys Ile Gly
Lys Asp Thr Cys Lys Glu Gly Glu Glu Asn Gln Lys 420
425 430 Ser Val Ile Ala Met Asp Gly Gly Leu
Phe Glu His Tyr Thr Lys Phe 435 440
445 Arg Lys Thr Met Gln Asp Thr Met Asn Glu Leu Leu Gly Glu
Glu Ile 450 455 460
Ser Lys Asn Ile Ile Val Glu Leu Ser Asn Asp Gly Ser Gly Leu Gly 465
470 475 480 Ala Ala Leu Leu Ala
Ala Ser His Ser Gln Tyr Leu Glu Tyr Ile Glu 485
490 495 Ser Thr 431854DNAPopulus trichocarpa
43atggggaagg tggcagtagg agcggcggtt gtttgcgcgg cgacagtgtg tgcggcggcg
60gcgttggtgg tgaggcacag gatgagatgt tcagggaggt gggccagggc tatggcgata
120ctaagagagt ttgaggaaaa ttgtgggacc cctattggga agttaagaca ggtggctgat
180gctatgaccg ttgagatgca tgccggcctt gcatctgagg gtggtagtaa gctcaagatg
240ttaatcagct atgtagataa tcttccctcc ggagaagaga atgggttgtt ctatgcattg
300gaccttggcg gaacaaattt tcgagttata cgggtactgc tcggtgggag ggatggaggt
360gttgtcaaac aagagtttga ggaagtttca attcctccac acttgatgac tggatcttca
420gatgcactat ttggtttcat tgctacagcg cttgccaatt ttgttgccac agaaagtgaa
480ggtctgcatt gttcacctgg gagacaaagg gagctcggtt ttaccttctc atttccagtt
540aggcaaacat caatagcatc tggaaatctt ataaaatgga caaaaggatt ctccatagat
600gatgtggttg gagaagatgt ggtgggagaa ttaaccaaag ccatggaaag aattggactt
660gacatgcgcg tgtcagcttt ggtcaatgat acaattggaa cattagctgg aggtcgatac
720cacaacccag atgtaattgc tgctgtaata ttgggtactg gaacaaatgc agcatatgta
780gagcgagcac aagcaattcc taagtggcat ggtcttctac ccaaatctgg agaaatggtt
840atcaacatgg aatggggtaa tttccggtct tcgcaccttc cactaacaga atatgatcaa
900gacttggatg ttgagtcttt gaatcctggc gaacagattt ttgaaaagat tatttctggt
960atgtatttgg gagaaattgt acgcagagtt ctactgaaaa tggctgaaga ggctgccttt
1020tttggtgata ttgttccaca aaaactgaag attccattca tcttaaggac gcctcacatg
1080tctgcaatgc accatgatga atcttcagat ctgagagttg ttggaagcaa actaaaagat
1140attttagaga tacctcatac ctcgctgaaa atgaggaaag ctattgttga actatgtgac
1200attgttgcca ctcgtggtgc ccgcctatct gctgctggga ttgtaggcat catcaagaaa
1260ttgggtagag acactgtaaa ggatggtgaa aagcagaagt ctgtgatagc aatggatggt
1320gggttgtatg agcactactc taaatttagt acctgcatgg aaagcactct caaggagtta
1380ctgggagaag aagtttctga caacattgtc gttgagcagt ctaatgatgg ctcaggcatt
1440ggagctgctc tcctagcagc ctcgcattcc caatacctgg aggtcgaaga atcttgatga
1500taacccatta cattaagtag tgttacattt ttgttttttg tatattatct atccctctac
1560actcgacttc aaatcatctg gagcttttca acctagttct tatgacgatg ccagcgaaat
1620ccatattttg tggtagtaag ttgagcagta gatcaaatac tcttttgaaa agtgccatta
1680gtggtcccaa gcttcagtgt gggagtctgt cacactgctc ctctttactt ttctcagtca
1740tttacatgaa ttacacaata aagtggggga tgtaaaacaa ttatttgagt tgttacattt
1800gacggaatat tttttctatg gttccgatgg aaaataaagt gaatttttat gatc
185444498PRTPopulus trichocarpa 44Met Gly Lys Val Ala Val Gly Ala Ala Val
Val Cys Ala Ala Thr Val 1 5 10
15 Cys Ala Ala Ala Ala Leu Val Val Arg His Arg Met Arg Cys Ser
Gly 20 25 30 Arg
Trp Ala Arg Ala Met Ala Ile Leu Arg Glu Phe Glu Glu Asn Cys 35
40 45 Gly Thr Pro Ile Gly Lys
Leu Arg Gln Val Ala Asp Ala Met Thr Val 50 55
60 Glu Met His Ala Gly Leu Ala Ser Glu Gly Gly
Ser Lys Leu Lys Met 65 70 75
80 Leu Ile Ser Tyr Val Asp Asn Leu Pro Ser Gly Glu Glu Asn Gly Leu
85 90 95 Phe Tyr
Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Ile Arg Val 100
105 110 Leu Leu Gly Gly Arg Asp Gly
Gly Val Val Lys Gln Glu Phe Glu Glu 115 120
125 Val Ser Ile Pro Pro His Leu Met Thr Gly Ser Ser
Asp Ala Leu Phe 130 135 140
Gly Phe Ile Ala Thr Ala Leu Ala Asn Phe Val Ala Thr Glu Ser Glu 145
150 155 160 Gly Leu His
Cys Ser Pro Gly Arg Gln Arg Glu Leu Gly Phe Thr Phe 165
170 175 Ser Phe Pro Val Arg Gln Thr Ser
Ile Ala Ser Gly Asn Leu Ile Lys 180 185
190 Trp Thr Lys Gly Phe Ser Ile Asp Asp Val Val Gly Glu
Asp Val Val 195 200 205
Gly Glu Leu Thr Lys Ala Met Glu Arg Ile Gly Leu Asp Met Arg Val 210
215 220 Ser Ala Leu Val
Asn Asp Thr Ile Gly Thr Leu Ala Gly Gly Arg Tyr 225 230
235 240 His Asn Pro Asp Val Ile Ala Ala Val
Ile Leu Gly Thr Gly Thr Asn 245 250
255 Ala Ala Tyr Val Glu Arg Ala Gln Ala Ile Pro Lys Trp His
Gly Leu 260 265 270
Leu Pro Lys Ser Gly Glu Met Val Ile Asn Met Glu Trp Gly Asn Phe
275 280 285 Arg Ser Ser His
Leu Pro Leu Thr Glu Tyr Asp Gln Asp Leu Asp Val 290
295 300 Glu Ser Leu Asn Pro Gly Glu Gln
Ile Phe Glu Lys Ile Ile Ser Gly 305 310
315 320 Met Tyr Leu Gly Glu Ile Val Arg Arg Val Leu Leu
Lys Met Ala Glu 325 330
335 Glu Ala Ala Phe Phe Gly Asp Ile Val Pro Gln Lys Leu Lys Ile Pro
340 345 350 Phe Ile Leu
Arg Thr Pro His Met Ser Ala Met His His Asp Glu Ser 355
360 365 Ser Asp Leu Arg Val Val Gly Ser
Lys Leu Lys Asp Ile Leu Glu Ile 370 375
380 Pro His Thr Ser Leu Lys Met Arg Lys Ala Ile Val Glu
Leu Cys Asp 385 390 395
400 Ile Val Ala Thr Arg Gly Ala Arg Leu Ser Ala Ala Gly Ile Val Gly
405 410 415 Ile Ile Lys Lys
Leu Gly Arg Asp Thr Val Lys Asp Gly Glu Lys Gln 420
425 430 Lys Ser Val Ile Ala Met Asp Gly Gly
Leu Tyr Glu His Tyr Ser Lys 435 440
445 Phe Ser Thr Cys Met Glu Ser Thr Leu Lys Glu Leu Leu Gly
Glu Glu 450 455 460
Val Ser Asp Asn Ile Val Val Glu Gln Ser Asn Asp Gly Ser Gly Ile 465
470 475 480 Gly Ala Ala Leu Leu
Ala Ala Ser His Ser Gln Tyr Leu Glu Val Glu 485
490 495 Glu Ser 451971DNASpinacia oleracea
45acgagcctta attagaccac ctcaaggcaa agctccgatt aaaccggtcc ctccaaaaaa
60aaaactcatc tgaattttcc ccaatcaaat tctcccggat ctactccttc tctcccttct
120ttctctctct agattgcatc ttgtaaagaa agaaagaaac caaaatttcc caacatgcca
180aattaatcca tcttctttcc attgttgtaa ttagagagag aaaatcaaat tgaagagcac
240ccatctgatt ttttttagtg gacgttacat agagagaaaa tgaggaaggc agcggtcgga
300gcagcggtgg tatgtacagc ggcggtgtgt gcggcggcag cggtgttggt aagacagagg
360atgaagagct caagcaagtg gggtcgtgta atggcaatac tgaaggagtt agatgacaat
420tgtgggaccc ctttgggtaa gcttagacag gtggctgatg ctatgaccgt tgagatgcac
480gctggtcttg catctgaggg tgcttctaag ctcaagatgc tcatcagcta cgttgacaat
540ctccccactg gggacgagca tggactgttc tatgcgttag accttggcgg caccaacttc
600cgagtgcttc gagtaaagtt gggtggcaaa gaaaaacgtg ttgtcgaaca agaatttgac
660gaagtatcaa ttccacctga gttgatggtt ggtacatcag aacaactgtt tgattatatc
720gccgaagccc tagccaaatt tgttgcaacg gaaagtgaag gtcttcatcc tgaacccaat
780aaacagagag agctgggatt taccttctct ttccctgtca agcagacatc aatcgcatca
840gggactctta taagatggac caagggcttt aatatagaag atacagttgg tgaagatgtg
900gtggctgaat tgaccaaggc catgctaaga aaaggtgtag atatgcgtgt gacagctttg
960gtcaacgata cagttggaac cctagctgga ggtaggtact ataaagaaga tgtaattgct
1020gctgttatat tgggtactgg aacaaacgca gcttacgttg aacgtgctag tgcaattcac
1080aagtggcatg gtcctttgcc caaatcaggg gagatggtaa ttaacatgga gtggggtaat
1140ttccgttcat cgtatttacc tttgactgaa tatgacatag cactagatga agaaagtttg
1200aatcctggtg aacagatatt tgagaaaatg atatcaggaa tgtacttggg tgagattgtg
1260cgaagagtct tgtataggat ggcagacgag gccagccttt ttggtgatac agtcccatca
1320aaattgaaaa ctccattcat cttaaggaca ccagacatgt ctgccatgca tcatgacaca
1380tcacctgatc ttaaagttgt tgcgagcaaa ctgaaggatg tccttgggat accaaactca
1440tcattaaagg tgcgaaagat tatagttgac gtatgtgacg tcattgcttc acgtggggcc
1500tgcatttctg cagccgggat cttgggtatt attaagaaac tagggagaga cacattgaag
1560caaggtgaga accagaagtc tgtgattgca ttagatggag ggttgtttga gcactacgcc
1620aaattccggg agtgcatgga ggactctttg aaggagctcc taggcgatga agtcgctgaa
1680actattgtaa ttgagcactc aaatgatgga tcaggcattg gtgctgctct tctagcagcg
1740tcgcattccc agtacctcga ggaagatgaa tcttgatgac aagatcctac atcctcaatt
1800ttgtataact ttcttcaagc tccttagatc cttagagatc atggaaattt tttctttttt
1860ttttcttttt tctttttaga ctttgctgcc ccagtgaaga ataaattatg gccaaacggg
1920ctctgcttgc tgcatgaaaa aatggcatga agctcgctgt tctctcgtgc c
197146498PRTSpinacia oleracea 46Met Arg Lys Ala Ala Val Gly Ala Ala Val
Val Cys Thr Ala Ala Val 1 5 10
15 Cys Ala Ala Ala Ala Val Leu Val Arg Gln Arg Met Lys Ser Ser
Ser 20 25 30 Lys
Trp Gly Arg Val Met Ala Ile Leu Lys Glu Leu Asp Asp Asn Cys 35
40 45 Gly Thr Pro Leu Gly Lys
Leu Arg Gln Val Ala Asp Ala Met Thr Val 50 55
60 Glu Met His Ala Gly Leu Ala Ser Glu Gly Ala
Ser Lys Leu Lys Met 65 70 75
80 Leu Ile Ser Tyr Val Asp Asn Leu Pro Thr Gly Asp Glu His Gly Leu
85 90 95 Phe Tyr
Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Arg Val 100
105 110 Lys Leu Gly Gly Lys Glu Lys
Arg Val Val Glu Gln Glu Phe Asp Glu 115 120
125 Val Ser Ile Pro Pro Glu Leu Met Val Gly Thr Ser
Glu Gln Leu Phe 130 135 140
Asp Tyr Ile Ala Glu Ala Leu Ala Lys Phe Val Ala Thr Glu Ser Glu 145
150 155 160 Gly Leu His
Pro Glu Pro Asn Lys Gln Arg Glu Leu Gly Phe Thr Phe 165
170 175 Ser Phe Pro Val Lys Gln Thr Ser
Ile Ala Ser Gly Thr Leu Ile Arg 180 185
190 Trp Thr Lys Gly Phe Asn Ile Glu Asp Thr Val Gly Glu
Asp Val Val 195 200 205
Ala Glu Leu Thr Lys Ala Met Leu Arg Lys Gly Val Asp Met Arg Val 210
215 220 Thr Ala Leu Val
Asn Asp Thr Val Gly Thr Leu Ala Gly Gly Arg Tyr 225 230
235 240 Tyr Lys Glu Asp Val Ile Ala Ala Val
Ile Leu Gly Thr Gly Thr Asn 245 250
255 Ala Ala Tyr Val Glu Arg Ala Ser Ala Ile His Lys Trp His
Gly Pro 260 265 270
Leu Pro Lys Ser Gly Glu Met Val Ile Asn Met Glu Trp Gly Asn Phe
275 280 285 Arg Ser Ser Tyr
Leu Pro Leu Thr Glu Tyr Asp Ile Ala Leu Asp Glu 290
295 300 Glu Ser Leu Asn Pro Gly Glu Gln
Ile Phe Glu Lys Met Ile Ser Gly 305 310
315 320 Met Tyr Leu Gly Glu Ile Val Arg Arg Val Leu Tyr
Arg Met Ala Asp 325 330
335 Glu Ala Ser Leu Phe Gly Asp Thr Val Pro Ser Lys Leu Lys Thr Pro
340 345 350 Phe Ile Leu
Arg Thr Pro Asp Met Ser Ala Met His His Asp Thr Ser 355
360 365 Pro Asp Leu Lys Val Val Ala Ser
Lys Leu Lys Asp Val Leu Gly Ile 370 375
380 Pro Asn Ser Ser Leu Lys Val Arg Lys Ile Ile Val Asp
Val Cys Asp 385 390 395
400 Val Ile Ala Ser Arg Gly Ala Cys Ile Ser Ala Ala Gly Ile Leu Gly
405 410 415 Ile Ile Lys Lys
Leu Gly Arg Asp Thr Leu Lys Gln Gly Glu Asn Gln 420
425 430 Lys Ser Val Ile Ala Leu Asp Gly Gly
Leu Phe Glu His Tyr Ala Lys 435 440
445 Phe Arg Glu Cys Met Glu Asp Ser Leu Lys Glu Leu Leu Gly
Asp Glu 450 455 460
Val Ala Glu Thr Ile Val Ile Glu His Ser Asn Asp Gly Ser Gly Ile 465
470 475 480 Gly Ala Ala Leu Leu
Ala Ala Ser His Ser Gln Tyr Leu Glu Glu Asp 485
490 495 Glu Ser 471626DNAOryza sativa
47ggttcaaagc ttgttcgatt cgttcgcgcg ggaccatggc ggcggcggcg gtggcggcag
60atcagaaggt ggtgacgatg acgagcctcc gggagggctg cgcttgcgcg gcgcctcctg
120ctgcagctgc gccgccgatg ccgaagatgg cggcggcgca gagggtggtg gcggagctga
180gagaagcgtg cgcgacgccg gcggcgaggc tggcggaggt ggccgcggcg atggccggcg
240agatggaggc cgggctggcg gtggagggcg gcagcagcga gatgaagatg atcgtgtcgt
300acgtcgacag cctccccacc ggcggcgagg aggggtcgta ctacgcgctc gacctcggcg
360gcaccaactt ccgcgtcctc cgcgtgcggc ttgccggcgg cggcgtcgcc gagcgcgtgg
420cgagggaggt cccgatccct cccggcctca tgtccggcgg cggcgccacc tcggagtgcc
480tcttcggctt catcgcctcc gcgctagccg agttcgtcgg cgaggaggaa gaagaaggcg
540gcctcgacgg cggcgagagg gagcttgggt tcaccttctc cttccccgtg caccaaacct
600ccatcgcgtc ggggacgctc atccggtgga cgaaggcgtt cgccgtcgac gacgcgatcg
660gcgaggacgt cgtggcggcg ctgcaggcgg ccatgtcgga gcgggggctc gacatgcgcg
720tgtcggcgct catcaacgac accgtcggga cgctcgccgc gggcagctac tacgacgagg
780acgtcgtggc cgccgtcatc ctcggcaccg gcacgaacgc cgcctacgtc gaggacgcca
840ccgccatcgc caagctacac ccatcgcagc tgccagcatc gaacaccatg gtgatcaaca
900ccgagtgggg cagcttcgcc tcgccgtgcc tcccattgac ggagttcgac gaagcactcg
960atcaggagag cctcaacccc ggcgagcaga cctacgagaa gctcatctcc gggatgtacc
1020tcggcgagat cgtcaggagg gtcctcctca agatctcctc ccggtgcccc tccctcctcg
1080gcggcgccgg cgagctcgcg acgccgttcg tcctcaggac acccgacgtg tccgcgatgc
1140accacgacga gacgcccgac ctgagcatcg tcggcgagaa gctggaacgc acgctgggca
1200tccgcggcac gtcgccggag gcgaggagga tggtcgtcga ggtgtgcgac atcgtcgcca
1260cgagggccgc ccggctggcc gcggcgggga tcgtcgggat cctgaagaag atcgggaggg
1320tcgacggcgg cgaggggcgg aggaggaggt cggtggtcgc cgtggacggc gggctgttcg
1380agcactacgg caagttccgg cggtgcatgg agagcgcggt gagggagctg ctcggagagg
1440cggcggcgga gagggtggtc gtcaagctcg ccagcgacgg ctccgggctc ggcgccgccc
1500tggttgcagc tgctcactcg cagagagcat aatattgaat tgtggtgaaa tatgcgtgtg
1560tttgaaacaa gcttgaatga tttgaatatt tgatgcgatt tgcaaaacca catatgaaat
1620agaagc
162648498PRTOryza sativa 48Met Ala Ala Ala Ala Val Ala Ala Asp Gln Lys
Val Val Thr Met Thr 1 5 10
15 Ser Leu Arg Glu Gly Cys Ala Cys Ala Ala Pro Pro Ala Ala Ala Ala
20 25 30 Pro Pro
Met Pro Lys Met Ala Ala Ala Gln Arg Val Val Ala Glu Leu 35
40 45 Arg Glu Ala Cys Ala Thr Pro
Ala Ala Arg Leu Ala Glu Val Ala Ala 50 55
60 Ala Met Ala Gly Glu Met Glu Ala Gly Leu Ala Val
Glu Gly Gly Ser 65 70 75
80 Ser Glu Met Lys Met Ile Val Ser Tyr Val Asp Ser Leu Pro Thr Gly
85 90 95 Gly Glu Glu
Gly Ser Tyr Tyr Ala Leu Asp Leu Gly Gly Thr Asn Phe 100
105 110 Arg Val Leu Arg Val Arg Leu Ala
Gly Gly Gly Val Ala Glu Arg Val 115 120
125 Ala Arg Glu Val Pro Ile Pro Pro Gly Leu Met Ser Gly
Gly Gly Ala 130 135 140
Thr Ser Glu Cys Leu Phe Gly Phe Ile Ala Ser Ala Leu Ala Glu Phe 145
150 155 160 Val Gly Glu Glu
Glu Glu Glu Gly Gly Leu Asp Gly Gly Glu Arg Glu 165
170 175 Leu Gly Phe Thr Phe Ser Phe Pro Val
His Gln Thr Ser Ile Ala Ser 180 185
190 Gly Thr Leu Ile Arg Trp Thr Lys Ala Phe Ala Val Asp Asp
Ala Ile 195 200 205
Gly Glu Asp Val Val Ala Ala Leu Gln Ala Ala Met Ser Glu Arg Gly 210
215 220 Leu Asp Met Arg Val
Ser Ala Leu Ile Asn Asp Thr Val Gly Thr Leu 225 230
235 240 Ala Ala Gly Ser Tyr Tyr Asp Glu Asp Val
Val Ala Ala Val Ile Leu 245 250
255 Gly Thr Gly Thr Asn Ala Ala Tyr Val Glu Asp Ala Thr Ala Ile
Ala 260 265 270 Lys
Leu His Pro Ser Gln Leu Pro Ala Ser Asn Thr Met Val Ile Asn 275
280 285 Thr Glu Trp Gly Ser Phe
Ala Ser Pro Cys Leu Pro Leu Thr Glu Phe 290 295
300 Asp Glu Ala Leu Asp Gln Glu Ser Leu Asn Pro
Gly Glu Gln Thr Tyr 305 310 315
320 Glu Lys Leu Ile Ser Gly Met Tyr Leu Gly Glu Ile Val Arg Arg Val
325 330 335 Leu Leu
Lys Ile Ser Ser Arg Cys Pro Ser Leu Leu Gly Gly Ala Gly 340
345 350 Glu Leu Ala Thr Pro Phe Val
Leu Arg Thr Pro Asp Val Ser Ala Met 355 360
365 His His Asp Glu Thr Pro Asp Leu Ser Ile Val Gly
Glu Lys Leu Glu 370 375 380
Arg Thr Leu Gly Ile Arg Gly Thr Ser Pro Glu Ala Arg Arg Met Val 385
390 395 400 Val Glu Val
Cys Asp Ile Val Ala Thr Arg Ala Ala Arg Leu Ala Ala 405
410 415 Ala Gly Ile Val Gly Ile Leu Lys
Lys Ile Gly Arg Val Asp Gly Gly 420 425
430 Glu Gly Arg Arg Arg Arg Ser Val Val Ala Val Asp Gly
Gly Leu Phe 435 440 445
Glu His Tyr Gly Lys Phe Arg Arg Cys Met Glu Ser Ala Val Arg Glu 450
455 460 Leu Leu Gly Glu
Ala Ala Ala Glu Arg Val Val Val Lys Leu Ala Ser 465 470
475 480 Asp Gly Ser Gly Leu Gly Ala Ala Leu
Val Ala Ala Ala His Ser Gln 485 490
495 Arg Ala 491741DNAOryza sativa 49atatgtacgt aagggcccca
tctcgccgag gagaggctgg gcgaccgcga tgaggaaggc 60ggcggcggcg gcggtcgcgg
cagcggcggc ggtcggcgtg gcgctgctgg tgcgccggca 120gctgcgggag gcgaagcggt
ggggccgggc cgacgcggtg ctgcgggagc tggaggagcg 180gtgcgcggcg ccgccggggc
ggctgcggca ggtggcggac gcgatggccg tcgagatgca 240cgccgggctc gcctccgagg
gcgggagcaa actcaagatg atcatcagct acgtcgacgc 300cctcccttcc ggggaagaga
agggggtgtt ttatgcgctt gaccttggag gtacaaattt 360ccgtgtttta cgggttcaat
taggtggcaa ggaagggaga gttatcaagc aagaacatga 420cgagatttca attcctccgc
atctgatgac tggtggttca aatgaactat ttgattttat 480tgcttcttct ttagcaaaat
ttgttgcttc agagggtgaa gactttcatc ttgctgaggg 540gaggcagaga gaacttggct
ttacgttctc tttcccagta aagcaaactt caattgcatc 600tggcactctt attaattgga
ctaagggttt ttcgattgat gaaacggttg gtgaagatgt 660tgtgactgaa ttaaccaagg
ctcttgaacg ccaggggctt gatatgaaag tcacagcatt 720gataaatgat actataggga
cattggctgg tgggagatat gatgataatg atgtcattgc 780tgctgttata ctgggaacag
gtactaatgc agcatacgtg gaacgtgcca atgcaattcc 840taaatggcat gacctcctgc
cgaagtcagg agatatggta ataaatatgg aatgggggaa 900cttcaggtca tcccatcttc
ctttgactga atttgatcaa gcattagatg ctgaaagttt 960gaatcctggt gaacaggttt
acgaaaagtt gatctctggc atgtatttgg gagaaattgt 1020tcgtagagtc ctattaaaga
tggctgaaga agcttctctt tttggtgatg aagtaccacc 1080aaaactcaag attccattta
ttatcaggac tccatacatg tcaatgatgc actgcgacag 1140atcacctgat ctcagaacag
ttggagcgaa actgaaagat atcctggggg tccaaaacac 1200ctcccttaaa acaagaaggc
ttgtggtgga tgtctgcgac atcgttgcga aacgcgccgc 1260tcaccttgct gctgcaggga
tacacgggat cctgaagaag cttgggcgcg acgtccccaa 1320caccgacaag cagaggacgg
tgatcgccgt cgacggtggg ctctacgagc actacaccat 1380cttcgccgag tgcgtggaga
gcaccctgag ggacatgctt ggggaggatg tgtcctccac 1440cattgtcatc aagctcgcca
aggacgggtc aggcattggc gctgctctcc ttgctgcggc 1500tcattctcaa taccgtgagg
ctgaggagct ctagtggatc tcttctcttg gcagctgtta 1560tgctttagtt gtttggattt
tcctgagatt ctgaaccccc aggagcaatt gcattctata 1620gagttgcacc agcaccgtgc
tggtgtttag aactttagat ggaggcacca acctttagta 1680gcttgcaagt cctgtaaact
gtataaacgc agaacacagt attgtccttc caacaaattt 1740c
174150494PRTOryza sativa
50Met Arg Lys Ala Ala Ala Ala Ala Val Ala Ala Ala Ala Ala Val Gly 1
5 10 15 Val Ala Leu Leu
Val Arg Arg Gln Leu Arg Glu Ala Lys Arg Trp Gly 20
25 30 Arg Ala Asp Ala Val Leu Arg Glu Leu
Glu Glu Arg Cys Ala Ala Pro 35 40
45 Pro Gly Arg Leu Arg Gln Val Ala Asp Ala Met Ala Val Glu
Met His 50 55 60
Ala Gly Leu Ala Ser Glu Gly Gly Ser Lys Leu Lys Met Ile Ile Ser 65
70 75 80 Tyr Val Asp Ala Leu
Pro Ser Gly Glu Glu Lys Gly Val Phe Tyr Ala 85
90 95 Leu Asp Leu Gly Gly Thr Asn Phe Arg Val
Leu Arg Val Gln Leu Gly 100 105
110 Gly Lys Glu Gly Arg Val Ile Lys Gln Glu His Asp Glu Ile Ser
Ile 115 120 125 Pro
Pro His Leu Met Thr Gly Gly Ser Asn Glu Leu Phe Asp Phe Ile 130
135 140 Ala Ser Ser Leu Ala Lys
Phe Val Ala Ser Glu Gly Glu Asp Phe His 145 150
155 160 Leu Ala Glu Gly Arg Gln Arg Glu Leu Gly Phe
Thr Phe Ser Phe Pro 165 170
175 Val Lys Gln Thr Ser Ile Ala Ser Gly Thr Leu Ile Asn Trp Thr Lys
180 185 190 Gly Phe
Ser Ile Asp Glu Thr Val Gly Glu Asp Val Val Thr Glu Leu 195
200 205 Thr Lys Ala Leu Glu Arg Gln
Gly Leu Asp Met Lys Val Thr Ala Leu 210 215
220 Ile Asn Asp Thr Ile Gly Thr Leu Ala Gly Gly Arg
Tyr Asp Asp Asn 225 230 235
240 Asp Val Ile Ala Ala Val Ile Leu Gly Thr Gly Thr Asn Ala Ala Tyr
245 250 255 Val Glu Arg
Ala Asn Ala Ile Pro Lys Trp His Asp Leu Leu Pro Lys 260
265 270 Ser Gly Asp Met Val Ile Asn Met
Glu Trp Gly Asn Phe Arg Ser Ser 275 280
285 His Leu Pro Leu Thr Glu Phe Asp Gln Ala Leu Asp Ala
Glu Ser Leu 290 295 300
Asn Pro Gly Glu Gln Val Tyr Glu Lys Leu Ile Ser Gly Met Tyr Leu 305
310 315 320 Gly Glu Ile Val
Arg Arg Val Leu Leu Lys Met Ala Glu Glu Ala Ser 325
330 335 Leu Phe Gly Asp Glu Val Pro Pro Lys
Leu Lys Ile Pro Phe Ile Ile 340 345
350 Arg Thr Pro Tyr Met Ser Met Met His Cys Asp Arg Ser Pro
Asp Leu 355 360 365
Arg Thr Val Gly Ala Lys Leu Lys Asp Ile Leu Gly Val Gln Asn Thr 370
375 380 Ser Leu Lys Thr Arg
Arg Leu Val Val Asp Val Cys Asp Ile Val Ala 385 390
395 400 Lys Arg Ala Ala His Leu Ala Ala Ala Gly
Ile His Gly Ile Leu Lys 405 410
415 Lys Leu Gly Arg Asp Val Pro Asn Thr Asp Lys Gln Arg Thr Val
Ile 420 425 430 Ala
Val Asp Gly Gly Leu Tyr Glu His Tyr Thr Ile Phe Ala Glu Cys 435
440 445 Val Glu Ser Thr Leu Arg
Asp Met Leu Gly Glu Asp Val Ser Ser Thr 450 455
460 Ile Val Ile Lys Leu Ala Lys Asp Gly Ser Gly
Ile Gly Ala Ala Leu 465 470 475
480 Leu Ala Ala Ala His Ser Gln Tyr Arg Glu Ala Glu Glu Leu
485 490 511692DNAOryza sativa
51tgggattcgt gggtgggttt gggagggagc ggctgcgggg cgggcggggg ggatggcgtg
60atcgcccgcg cgctctggag gggagggggg agggagaggg ggaggggtgg aggaggagat
120ggggagggtg gggctcggcg tggcggtggg gtgcgcggcg gtgacctgcg cgatcgccgc
180ggcgctcgtg gcgcgcaggg cgtcggcgcg ggcgcggtgg cggcgggcgg tggcgctgct
240gcgggagttc gaggaggggt gtgccacgcc gcccgcgagg ctgcgccagg tcgtggacgc
300catggtcgtc gagatgcacg ccggcctcgc gtccgatggc ggcagcaagc tcaagatgct
360gctcaccttc gtcgacgcgc tccccagcgg gagtgaagaa ggtgtatatt attctattga
420tcttggagga acaaacttca gagtcttgag ggtacaagtt ggtgcgggat ctgtgatcgt
480caaccaaaag gttgaacagc aacccatccc tgaggaactg accaaaggca ctactgaggg
540tttattcaac tttgttgccc tggcactaaa gaattttctt gaaggagaag atgaccaaga
600tggaaaaatg gcacttggtt ttacattttc tttccctgtt agacaaattt cagtgtcttc
660agggtcatta attaggtgga caaaaggatt ttccatcaga gacacggttg gcagagatgt
720tgctcagtgc ttaaatgaag cgcttgccaa ttgtgggcta aatgtgcggg tcactgcatt
780ggtgaatgat acagtgggga cattagctct agggcattac tatgatgaag acacagtggc
840tgctgtgata attgggtctg gcaccaacgc ttgctacatt gaacgcactg atgcaattat
900caagtgccag ggtcttctaa cgaactctgg aggcatggta gtaaacatgg agtgggggaa
960tttctggtca tcacatttgc caaggacgcc atatgacatc ttgctggatg atgaaacaca
1020caatcgcaat gatcagggct ttgagaaaat gatatcagga atgtatcttg gggaaattgc
1080aagattggta tttcatagaa tggcccagga atcagatgtt tttggtgatg ctgctgatag
1140tctatccaac cctttcattt tgagcacacc gtttctggcc gcaattcgcg aggacgattc
1200accagatctg agcgaagtca gaaggatact tcgagaacat ctgaagattc ccgatgctcc
1260tctgaaaact cgacggctgg tcgtgaaagt ctgcgacatt gtgactcgca gagccgcccg
1320tctagccgct gcaggcatcg tggggatact gaagaagctg gggagggatg ggagcggcgc
1380ggcgtcgagc gggagaggta gagggcagcc gaggaggacg gtggtggcga tcgagggcgg
1440gctgtaccag ggttacccgg tgttcaggga gtacctggac gaggccctgg tggagatcct
1500gggggaggag gtggcgcgga acgtgacgct gagggtgacg gaggatgggt cgggggtcgg
1560ggctgccctc ctcgccgccg tacattcgtc gaatagacag caacaaggag gtcccatata
1620gtgagagaga caatgaagat acagctagcc cctcttgttc aaatgtaaaa aagggacatt
1680gtttgatatc ta
169252500PRTOryza sativa 52Met Gly Arg Val Gly Leu Gly Val Ala Val Gly
Cys Ala Ala Val Thr 1 5 10
15 Cys Ala Ile Ala Ala Ala Leu Val Ala Arg Arg Ala Ser Ala Arg Ala
20 25 30 Arg Trp
Arg Arg Ala Val Ala Leu Leu Arg Glu Phe Glu Glu Gly Cys 35
40 45 Ala Thr Pro Pro Ala Arg Leu
Arg Gln Val Val Asp Ala Met Val Val 50 55
60 Glu Met His Ala Gly Leu Ala Ser Asp Gly Gly Ser
Lys Leu Lys Met 65 70 75
80 Leu Leu Thr Phe Val Asp Ala Leu Pro Ser Gly Ser Glu Glu Gly Val
85 90 95 Tyr Tyr Ser
Ile Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Arg Val 100
105 110 Gln Val Gly Ala Gly Ser Val Ile
Val Asn Gln Lys Val Glu Gln Gln 115 120
125 Pro Ile Pro Glu Glu Leu Thr Lys Gly Thr Thr Glu Gly
Leu Phe Asn 130 135 140
Phe Val Ala Leu Ala Leu Lys Asn Phe Leu Glu Gly Glu Asp Asp Gln 145
150 155 160 Asp Gly Lys Met
Ala Leu Gly Phe Thr Phe Ser Phe Pro Val Arg Gln 165
170 175 Ile Ser Val Ser Ser Gly Ser Leu Ile
Arg Trp Thr Lys Gly Phe Ser 180 185
190 Ile Arg Asp Thr Val Gly Arg Asp Val Ala Gln Cys Leu Asn
Glu Ala 195 200 205
Leu Ala Asn Cys Gly Leu Asn Val Arg Val Thr Ala Leu Val Asn Asp 210
215 220 Thr Val Gly Thr Leu
Ala Leu Gly His Tyr Tyr Asp Glu Asp Thr Val 225 230
235 240 Ala Ala Val Ile Ile Gly Ser Gly Thr Asn
Ala Cys Tyr Ile Glu Arg 245 250
255 Thr Asp Ala Ile Ile Lys Cys Gln Gly Leu Leu Thr Asn Ser Gly
Gly 260 265 270 Met
Val Val Asn Met Glu Trp Gly Asn Phe Trp Ser Ser His Leu Pro 275
280 285 Arg Thr Pro Tyr Asp Ile
Leu Leu Asp Asp Glu Thr His Asn Arg Asn 290 295
300 Asp Gln Gly Phe Glu Lys Met Ile Ser Gly Met
Tyr Leu Gly Glu Ile 305 310 315
320 Ala Arg Leu Val Phe His Arg Met Ala Gln Glu Ser Asp Val Phe Gly
325 330 335 Asp Ala
Ala Asp Ser Leu Ser Asn Pro Phe Ile Leu Ser Thr Pro Phe 340
345 350 Leu Ala Ala Ile Arg Glu Asp
Asp Ser Pro Asp Leu Ser Glu Val Arg 355 360
365 Arg Ile Leu Arg Glu His Leu Lys Ile Pro Asp Ala
Pro Leu Lys Thr 370 375 380
Arg Arg Leu Val Val Lys Val Cys Asp Ile Val Thr Arg Arg Ala Ala 385
390 395 400 Arg Leu Ala
Ala Ala Gly Ile Val Gly Ile Leu Lys Lys Leu Gly Arg 405
410 415 Asp Gly Ser Gly Ala Ala Ser Ser
Gly Arg Gly Arg Gly Gln Pro Arg 420 425
430 Arg Thr Val Val Ala Ile Glu Gly Gly Leu Tyr Gln Gly
Tyr Pro Val 435 440 445
Phe Arg Glu Tyr Leu Asp Glu Ala Leu Val Glu Ile Leu Gly Glu Glu 450
455 460 Val Ala Arg Asn
Val Thr Leu Arg Val Thr Glu Asp Gly Ser Gly Val 465 470
475 480 Gly Ala Ala Leu Leu Ala Ala Val His
Ser Ser Asn Arg Gln Gln Gln 485 490
495 Gly Gly Pro Ile 500 531937DNAOryza sativa
53tagtacgtgt agtgaggagc atttcttgcc acctttattc gattccacca cttgacacat
60ccatcgatca ctcctccccc taccaacgat cagctcagct tcagcttctc ctcggcatgt
120ccgccgccgc cgccatcgcg tcgccgatcc cggcggcgat cgccgtcgtg cagcagcaga
180ggcgggggag gagccgcggc ggcggctccg gcgctgccgc cgtccgctgc tccgcggtgg
240ccccgacgtc cgcgatcgcg cccatccttg ccgacctgag gctgcggtgc gccgccccgc
300tccccgtgct gcggcgcgtg gcggacgcca tggcctccgg gatgcgcgcc gggctggccg
360acgacggcgc cggcgagctc aagatgatcc ccagccacgt ctactcactc cccactggga
420atgaaacagg actgttttat gctctggacc ttggaggcac caactttaga gtgctgaggg
480tacaattggg aggaaaagat aagcgcatta tagataccga gtttgagcaa gtctcgatcc
540caagagaaat catgcatggt ataaccgagg atttgtttga tttcatcgcg agtggcctgt
600cgagatttgt agcaacggag ggtgataagt ttcatttgcc acaagggaga aagagagagt
660taggcttcac attctccttt ccggtgaatc agacttctat tgattctggc attctgatca
720agtggacaaa aggttttgct gtctctggaa ctgctgggaa agatgttgtt gcttgtctga
780atgctgcaat ggagaggcaa ggccttgata tgcgtgtctc tgccttggta aatgatactg
840tgggaacctt agctggagca cgttattggg acgatgacgt aatggtcgcg gtgattttgg
900gcactggcac caatgcatgc tacattcaac gaactgaagc tattccaaaa ttgcaacacc
960ttaagcttga aacaggaaac acgattatta acactgagtg gggagctttc tcagatggac
1020ttccattgac tgaatttgac agagaaatgg atgatgagag cataaatcct ggtgaacaga
1080tattcgagaa gacgatttcc gggatgtatc taggtgaaat tgtccgtagg gtgctggtca
1140agatggctga agtatctgat ctctttggtc attccttccc caagaaactt gctgaaccat
1200ttgttctaag gacaccacat ctatgcgcta tgcaacaaga cacctctgac aatcttggag
1260aagttgagtc catcttgagt gatgtcatcg gcgtgtccca agcttctctg ctggcacgga
1320gagtcactgt agaagtctcc gactgcatca tcaggagagg aggccggttg gccggggcag
1380gaatcgtagg gatccttgag aagatggaga atgactccag agggcacatt ttcggacgaa
1440gaacagtggt cgcgatggac ggtggtctat atgagaagta ccctcagtac aggaggtaca
1500tgaaggaggc tgtggctgag ctactgggac ccgagcgatc gaatcgtatc gccatcgagc
1560acacgaaaga cggatcaggg atcggcgctg cgctgttggc agctgcaaac tcaaagtatg
1620ctgctgctca aatctctaca aggtgatcgc aattgttata attgttgagt tactgggccg
1680aattaattct ctacaaggtg atctcaattg ttatcattgt tgagctactg ggacctgaac
1740aatccaagca tatcatcata tggtggcctc ttatctgaag tatggagggg cttcatatat
1800gctattcttc tgtactacta gtctactata ctatataacg catttgagga aaagaaacac
1860cagaaattta tcaacttatg ttttgtataa ttcatcaact tatgtgtttg tatagttaca
1920gttcaccaga aattcat
193754509PRTOryza sativa 54Met Ser Ala Ala Ala Ala Ile Ala Ser Pro Ile
Pro Ala Ala Ile Ala 1 5 10
15 Val Val Gln Gln Gln Arg Arg Gly Arg Ser Arg Gly Gly Gly Ser Gly
20 25 30 Ala Ala
Ala Val Arg Cys Ser Ala Val Ala Pro Thr Ser Ala Ile Ala 35
40 45 Pro Ile Leu Ala Asp Leu Arg
Leu Arg Cys Ala Ala Pro Leu Pro Val 50 55
60 Leu Arg Arg Val Ala Asp Ala Met Ala Ser Gly Met
Arg Ala Gly Leu 65 70 75
80 Ala Asp Asp Gly Ala Gly Glu Leu Lys Met Ile Pro Ser His Val Tyr
85 90 95 Ser Leu Pro
Thr Gly Asn Glu Thr Gly Leu Phe Tyr Ala Leu Asp Leu 100
105 110 Gly Gly Thr Asn Phe Arg Val Leu
Arg Val Gln Leu Gly Gly Lys Asp 115 120
125 Lys Arg Ile Ile Asp Thr Glu Phe Glu Gln Val Ser Ile
Pro Arg Glu 130 135 140
Ile Met His Gly Ile Thr Glu Asp Leu Phe Asp Phe Ile Ala Ser Gly 145
150 155 160 Leu Ser Arg Phe
Val Ala Thr Glu Gly Asp Lys Phe His Leu Pro Gln 165
170 175 Gly Arg Lys Arg Glu Leu Gly Phe Thr
Phe Ser Phe Pro Val Asn Gln 180 185
190 Thr Ser Ile Asp Ser Gly Ile Leu Ile Lys Trp Thr Lys Gly
Phe Ala 195 200 205
Val Ser Gly Thr Ala Gly Lys Asp Val Val Ala Cys Leu Asn Ala Ala 210
215 220 Met Glu Arg Gln Gly
Leu Asp Met Arg Val Ser Ala Leu Val Asn Asp 225 230
235 240 Thr Val Gly Thr Leu Ala Gly Ala Arg Tyr
Trp Asp Asp Asp Val Met 245 250
255 Val Ala Val Ile Leu Gly Thr Gly Thr Asn Ala Cys Tyr Ile Gln
Arg 260 265 270 Thr
Glu Ala Ile Pro Lys Leu Gln His Leu Lys Leu Glu Thr Gly Asn 275
280 285 Thr Ile Ile Asn Thr Glu
Trp Gly Ala Phe Ser Asp Gly Leu Pro Leu 290 295
300 Thr Glu Phe Asp Arg Glu Met Asp Asp Glu Ser
Ile Asn Pro Gly Glu 305 310 315
320 Gln Ile Phe Glu Lys Thr Ile Ser Gly Met Tyr Leu Gly Glu Ile Val
325 330 335 Arg Arg
Val Leu Val Lys Met Ala Glu Val Ser Asp Leu Phe Gly His 340
345 350 Ser Phe Pro Lys Lys Leu Ala
Glu Pro Phe Val Leu Arg Thr Pro His 355 360
365 Leu Cys Ala Met Gln Gln Asp Thr Ser Asp Asn Leu
Gly Glu Val Glu 370 375 380
Ser Ile Leu Ser Asp Val Ile Gly Val Ser Gln Ala Ser Leu Leu Ala 385
390 395 400 Arg Arg Val
Thr Val Glu Val Ser Asp Cys Ile Ile Arg Arg Gly Gly 405
410 415 Arg Leu Ala Gly Ala Gly Ile Val
Gly Ile Leu Glu Lys Met Glu Asn 420 425
430 Asp Ser Arg Gly His Ile Phe Gly Arg Arg Thr Val Val
Ala Met Asp 435 440 445
Gly Gly Leu Tyr Glu Lys Tyr Pro Gln Tyr Arg Arg Tyr Met Lys Glu 450
455 460 Ala Val Ala Glu
Leu Leu Gly Pro Glu Arg Ser Asn Arg Ile Ala Ile 465 470
475 480 Glu His Thr Lys Asp Gly Ser Gly Ile
Gly Ala Ala Leu Leu Ala Ala 485 490
495 Ala Asn Ser Lys Tyr Ala Ala Ala Gln Ile Ser Thr Arg
500 505 552058DNAOryza sativa
55ctctcgtcct cctttctcct acgcggccgg agagaggagg ggggaagaga ggatctcgca
60gagcgccata gcctcggatc cagaagcgga attcggtgga ggtctcgggc acctggatcg
120atcggaggaa gggaaggcgg agcagcggtg atggggaagg cggcggcggt ggggacggcg
180gtggtggtgg ccgcggcggt cggggtggcg gtggtgctgg cgcggaggcg gaggaggagg
240gacctggagc tggtggaggg agccgcggcg gagaggaaga ggaaggtggc ggcggtgatc
300gaggacgtgg agcacgcgct gtcgaccccg acggcgctgc tgcggggcat ctcggacgcc
360atggtcaccg agatggagcg aggcctgcgc ggggacagcc acgccatggt taagatgctc
420atcacctacg tcgacaacct ccccaccgga aatgaacagg ggttgtttta tgcattggat
480cttggaggaa ccaacttccg cgtcctgcga gtccaactcg gtggcaagga gaaacgtgtc
540gtccaacaac agtacgagga agtctcaatt ccaccacatc tgatggttgg aacttccatg
600gaactgtttg attttattgc ttctgcattg tcaaaatttg ttgatactga aggtgatgat
660ttccacctcc cagaagggag acagagagag ctgggcttca ccttttcctt cccagtgagc
720cagacatcaa tatcgtcagg aacgctcatc aagtggacaa agggtttctc catcaatgac
780gcggttggcg aagatgttgt atctgagttg ggcaaggcca tggagaggca gggattagat
840atgaaaattg cagcattggt taatgacact gtcggcacat tggctggtgg gaggtatgcg
900gataacagtg ttgttgctgc tataatattg ggcactggta caaatgcagc atatgttgag
960aatgctaatg caattcctaa atggaccggt ttactgccta ggtccggaaa tatggtaatc
1020aacacagaat gggggagctt taaatcagat aagcttcctc tttcagaatt cgataaagca
1080atggattttg aaagcttgaa tcctggagag cagatatatg aaaagttgat ttctggcatg
1140tatcttggag agatagtgcg aagaatcttg ctgaagcttg ctcatgatgc agctttgttt
1200ggggatgttg ttccatctaa gctagagcaa ccgtttgtac taaggacacc ggatatgtca
1260gccatgcatc atgactcgtc acatgacctt aaaactgttg gagctaagct aaaggatatc
1320gtcggggtcc cagatacttc cctggaagta agatacatta ccagtcacat ttgtgacata
1380gttgcagagc gtgctgcacg cttggctgct gctggcatat atggggtcct aaagaagcta
1440ggtcgggaca agatgccaaa agacggcagt aagatgccta ggactgtcat tgccttggat
1500ggtgggctct atgaacatta caagaagttc agcagttgct tagaatcaac tctaacagac
1560cttcttgggg atgatgtctc gtcttcggtg gttaccaagc tggccaacga tggttctggc
1620attggagctg ctcttctcgc agcctcgcac tcccagtatg ccgagatcga ctagctttaa
1680ggatgatctt gatgaatgat gaatcaaact ccgtttgtag gttctcattt cccccttcaa
1740aatccacata atactcctgg ctcccccctt gaaatcttac catctttttt tggctattct
1800gagggcaaac ataagtgcct ctgcagcggg atatagctag tatagcgcca atgagtttgg
1860aggttttcta atggcataaa acgttggatg gcagtagcag actaacaggg aaatggaggc
1920acaggcaatt tccattcctg ttctgtcaga ttcttttccc ccttaattga tgttgagaac
1980caagattttt ttgctctgta ttttctcttc gtaataaaga aggggacata atctaattgc
2040tcttgtttga tctcataa
205856507PRTOryza sativa 56Met Gly Lys Ala Ala Ala Val Gly Thr Ala Val
Val Val Ala Ala Ala 1 5 10
15 Val Gly Val Ala Val Val Leu Ala Arg Arg Arg Arg Arg Arg Asp Leu
20 25 30 Glu Leu
Val Glu Gly Ala Ala Ala Glu Arg Lys Arg Lys Val Ala Ala 35
40 45 Val Ile Glu Asp Val Glu His
Ala Leu Ser Thr Pro Thr Ala Leu Leu 50 55
60 Arg Gly Ile Ser Asp Ala Met Val Thr Glu Met Glu
Arg Gly Leu Arg 65 70 75
80 Gly Asp Ser His Ala Met Val Lys Met Leu Ile Thr Tyr Val Asp Asn
85 90 95 Leu Pro Thr
Gly Asn Glu Gln Gly Leu Phe Tyr Ala Leu Asp Leu Gly 100
105 110 Gly Thr Asn Phe Arg Val Leu Arg
Val Gln Leu Gly Gly Lys Glu Lys 115 120
125 Arg Val Val Gln Gln Gln Tyr Glu Glu Val Ser Ile Pro
Pro His Leu 130 135 140
Met Val Gly Thr Ser Met Glu Leu Phe Asp Phe Ile Ala Ser Ala Leu 145
150 155 160 Ser Lys Phe Val
Asp Thr Glu Gly Asp Asp Phe His Leu Pro Glu Gly 165
170 175 Arg Gln Arg Glu Leu Gly Phe Thr Phe
Ser Phe Pro Val Ser Gln Thr 180 185
190 Ser Ile Ser Ser Gly Thr Leu Ile Lys Trp Thr Lys Gly Phe
Ser Ile 195 200 205
Asn Asp Ala Val Gly Glu Asp Val Val Ser Glu Leu Gly Lys Ala Met 210
215 220 Glu Arg Gln Gly Leu
Asp Met Lys Ile Ala Ala Leu Val Asn Asp Thr 225 230
235 240 Val Gly Thr Leu Ala Gly Gly Arg Tyr Ala
Asp Asn Ser Val Val Ala 245 250
255 Ala Ile Ile Leu Gly Thr Gly Thr Asn Ala Ala Tyr Val Glu Asn
Ala 260 265 270 Asn
Ala Ile Pro Lys Trp Thr Gly Leu Leu Pro Arg Ser Gly Asn Met 275
280 285 Val Ile Asn Thr Glu Trp
Gly Ser Phe Lys Ser Asp Lys Leu Pro Leu 290 295
300 Ser Glu Phe Asp Lys Ala Met Asp Phe Glu Ser
Leu Asn Pro Gly Glu 305 310 315
320 Gln Ile Tyr Glu Lys Leu Ile Ser Gly Met Tyr Leu Gly Glu Ile Val
325 330 335 Arg Arg
Ile Leu Leu Lys Leu Ala His Asp Ala Ala Leu Phe Gly Asp 340
345 350 Val Val Pro Ser Lys Leu Glu
Gln Pro Phe Val Leu Arg Thr Pro Asp 355 360
365 Met Ser Ala Met His His Asp Ser Ser His Asp Leu
Lys Thr Val Gly 370 375 380
Ala Lys Leu Lys Asp Ile Val Gly Val Pro Asp Thr Ser Leu Glu Val 385
390 395 400 Arg Tyr Ile
Thr Ser His Ile Cys Asp Ile Val Ala Glu Arg Ala Ala 405
410 415 Arg Leu Ala Ala Ala Gly Ile Tyr
Gly Val Leu Lys Lys Leu Gly Arg 420 425
430 Asp Lys Met Pro Lys Asp Gly Ser Lys Met Pro Arg Thr
Val Ile Ala 435 440 445
Leu Asp Gly Gly Leu Tyr Glu His Tyr Lys Lys Phe Ser Ser Cys Leu 450
455 460 Glu Ser Thr Leu
Thr Asp Leu Leu Gly Asp Asp Val Ser Ser Ser Val 465 470
475 480 Val Thr Lys Leu Ala Asn Asp Gly Ser
Gly Ile Gly Ala Ala Leu Leu 485 490
495 Ala Ala Ser His Ser Gln Tyr Ala Glu Ile Asp
500 505 572070DNAOryza sativa 57gaggaaggag
gaggagtagg acgctgcagt ggtgggtggc gtagctcccg atccgggaag 60ccgacccggt
ctgggggatt tgctcctggc gcgcgctcga tcgagaggga ggccagggtt 120gggttggggg
gagcgtgaag aagcgcgcgc gcggctatgg ggaaggggac ggtagtgggg 180acggcggtgg
tggtgtgcgc tgcagcggcc gcggcggttg gggtggcggt ggtggtgtcg 240cggaggagga
ggagcaagcg ggaggcggag gaggagcggc ggaggagggc cgccgctgtg 300atcgaggagg
tggagcagag gttctcgacg cccacggcgc tgctgcgcgg catcgcggac 360gccatggtgg
aggagatgga gcgcggcctc cgcgccgacc ctcacgcccc gctcaagatg 420ctcatcagct
acgtcgacaa cctccccacc ggggatgagc acggactgtt ctatgctctc 480gatcttgggg
gcaccaattt ccgtgttata cgtgttcagc ttggaggaag ggaaaagcgt 540gttgttagtc
aacagtacga agaggttgcc attccacctc acctgatggt tgggacttct 600atggaactgt
ttgacttcat tgcggctgag ttggaaagtt ttgtcaagac cgagggagag 660gatttccact
tgccagaggg caggcagaga gagttaggct tcaccttttc tttcccagtg 720caccaaacat
caatatcatc aggcactctt attaagtgga caaagggatt ttccatcaat 780ggcacggtgg
gggaagatgt tgtggctgaa ttgagcaggg ctatggaaag gcaagggctt 840gatatgaaag
ttacagctct tgttaatgac actgtaggca cattggctgg cggaagatat 900gttgataatg
acgttgctgc tgctgtaata ttaggcactg gcacaaacgc agcctacgtg 960gagcatgcaa
atgcaattcc aaaatggact ggattactac ctagatcagg aaatatggtg 1020attaacatgg
aatggggaaa cttcaagtca gaaaggcttc ctcgttcaga ttacgataat 1080gccttggact
ttgaaagttt aaacccaggc gagcagatat atgaaaagat gatttccggc 1140atgtatcttg
gagagattgt gcgcagaatc ttgcttaagc ttgctcatga tgcttccttg 1200tttggagatg
ttgttccaac aaagctggag cagcgcttta tactgaggac gccggacatg 1260tcagcgatgc
atcatgatac ctcacatgat ctgaaacacc tgggagctaa gctgaaggat 1320atcctggggg
tcgctgatac ttccctggaa gcacgataca tcacccttca tgtctgcgac 1380ctcgttgcag
agagaggtgc acgcttagct gctgctggta tatatggcat tctaaagaag 1440ctgggcaggg
acagagtgcc aagtgacggt agtcaaaagc agaggactgt cattgctctg 1500gatggtggtc
tctatgagca ttacaagaag ttcagaacct gcctagaagc aacgcttgca 1560gacctgcttg
gagaggaggc tgcctcatca gttgttgtca agttggcaaa cgatggctct 1620ggcatcggag
ctgcacttct tgcagcatct cactcccagt atgctagcgt cgagtagtaa 1680caggagctca
tgggactgag ctcccagtgt agcttgtttt cctcccattt tccccgtttc 1740tttccaatgg
gagttcgttt ccctcctgcg attcgcatct ccttttgcta ttctgcagtc 1800acataaacga
gtgcctgtgc agcgggatgt agctagtatg gcgccaaaga gtttgcagtt 1860atcacatgaa
caagcatttg caactgcagg gaagtgaaaa cgggggcttg aatgatgccg 1920ttcttttcct
gcaaattatt ttcccccttt ccctgtaagt ttgtattgtg atgcgatgtc 1980gcaaaccaat
cacagcggtt tcgcgtgtag ccttttgtca ttcagatttc agaataaaga 2040gggggacata
atttcacttt tcgtatgtca
207058506PRTOryza sativa 58Met Gly Lys Gly Thr Val Val Gly Thr Ala Val
Val Val Cys Ala Ala 1 5 10
15 Ala Ala Ala Ala Val Gly Val Ala Val Val Val Ser Arg Arg Arg Arg
20 25 30 Ser Lys
Arg Glu Ala Glu Glu Glu Arg Arg Arg Arg Ala Ala Ala Val 35
40 45 Ile Glu Glu Val Glu Gln Arg
Phe Ser Thr Pro Thr Ala Leu Leu Arg 50 55
60 Gly Ile Ala Asp Ala Met Val Glu Glu Met Glu Arg
Gly Leu Arg Ala 65 70 75
80 Asp Pro His Ala Pro Leu Lys Met Leu Ile Ser Tyr Val Asp Asn Leu
85 90 95 Pro Thr Gly
Asp Glu His Gly Leu Phe Tyr Ala Leu Asp Leu Gly Gly 100
105 110 Thr Asn Phe Arg Val Ile Arg Val
Gln Leu Gly Gly Arg Glu Lys Arg 115 120
125 Val Val Ser Gln Gln Tyr Glu Glu Val Ala Ile Pro Pro
His Leu Met 130 135 140
Val Gly Thr Ser Met Glu Leu Phe Asp Phe Ile Ala Ala Glu Leu Glu 145
150 155 160 Ser Phe Val Lys
Thr Glu Gly Glu Asp Phe His Leu Pro Glu Gly Arg 165
170 175 Gln Arg Glu Leu Gly Phe Thr Phe Ser
Phe Pro Val His Gln Thr Ser 180 185
190 Ile Ser Ser Gly Thr Leu Ile Lys Trp Thr Lys Gly Phe Ser
Ile Asn 195 200 205
Gly Thr Val Gly Glu Asp Val Val Ala Glu Leu Ser Arg Ala Met Glu 210
215 220 Arg Gln Gly Leu Asp
Met Lys Val Thr Ala Leu Val Asn Asp Thr Val 225 230
235 240 Gly Thr Leu Ala Gly Gly Arg Tyr Val Asp
Asn Asp Val Ala Ala Ala 245 250
255 Val Ile Leu Gly Thr Gly Thr Asn Ala Ala Tyr Val Glu His Ala
Asn 260 265 270 Ala
Ile Pro Lys Trp Thr Gly Leu Leu Pro Arg Ser Gly Asn Met Val 275
280 285 Ile Asn Met Glu Trp Gly
Asn Phe Lys Ser Glu Arg Leu Pro Arg Ser 290 295
300 Asp Tyr Asp Asn Ala Leu Asp Phe Glu Ser Leu
Asn Pro Gly Glu Gln 305 310 315
320 Ile Tyr Glu Lys Met Ile Ser Gly Met Tyr Leu Gly Glu Ile Val Arg
325 330 335 Arg Ile
Leu Leu Lys Leu Ala His Asp Ala Ser Leu Phe Gly Asp Val 340
345 350 Val Pro Thr Lys Leu Glu Gln
Arg Phe Ile Leu Arg Thr Pro Asp Met 355 360
365 Ser Ala Met His His Asp Thr Ser His Asp Leu Lys
His Leu Gly Ala 370 375 380
Lys Leu Lys Asp Ile Leu Gly Val Ala Asp Thr Ser Leu Glu Ala Arg 385
390 395 400 Tyr Ile Thr
Leu His Val Cys Asp Leu Val Ala Glu Arg Gly Ala Arg 405
410 415 Leu Ala Ala Ala Gly Ile Tyr Gly
Ile Leu Lys Lys Leu Gly Arg Asp 420 425
430 Arg Val Pro Ser Asp Gly Ser Gln Lys Gln Arg Thr Val
Ile Ala Leu 435 440 445
Asp Gly Gly Leu Tyr Glu His Tyr Lys Lys Phe Arg Thr Cys Leu Glu 450
455 460 Ala Thr Leu Ala
Asp Leu Leu Gly Glu Glu Ala Ala Ser Ser Val Val 465 470
475 480 Val Lys Leu Ala Asn Asp Gly Ser Gly
Ile Gly Ala Ala Leu Leu Ala 485 490
495 Ala Ser His Ser Gln Tyr Ala Ser Val Glu 500
505 591816DNAOryza sativa 59ctttgatctt gaccaccaat
ctcgcatttc ttggatcttc tcgtcttcct cactcctcga 60tcgatccatc gattggttgc
tggtcggaga ccatggtggc ggcggcggtg gcggcggcgg 120agcaggtggt ggcggcgctg
cgggaggagt gcgcgacgcc ggcggcgcgg ctggatgggg 180tggcggcggc gatggccggc
gagatggcgg cggggttggc ggaggagggc ggcagcaaga 240tcaagatgat cgtttcctac
gtcgacaacc tccccaatgg gaccgaagag ggcttgttct 300acgcgctgga tcttggggga
acaaacttcc gagtcctgcg tgtgcagctt gctggaaagg 360agaaacgggt cgtgaagcgc
gagtcgaggg aggtctcgat ccctccccat ctcatgtccg 420gcaactcctc ggagctgttt
ggcttcatcg cttccgcgtt agccaagttc gtcgctgatg 480aaggacataa tgccgtgttc
aacgacaggc aaagggaact ggggttcacc ttctctttcc 540ctgtgcggca aacatcgatt
gcgtctggga ctcttatcaa gtggaccaag gcgttttcta 600ttgatgatgc tgtaggtgaa
gatgtggtgg ccgaattgca gatggccatg gagaagcaag 660gtctggacat gcgcgtttcc
gcattgacca atgacactgt tgggacattg gcggcaggca 720gctactacga cgaagatatt
gtcgtcggtg tgatcttagg cactggctca aacgccgcat 780atcttgagaa ggcaaatgct
atccctaagt tggaaggcga gttaccaaaa tcaggaaaca 840tggttattaa tacagaatgg
ggtaacttct cctcatcctg tcttccaata acagaatatg 900atgaagcact agataaagag
agcttaaacc ctggagagca gatcttcgag aaattgattt 960caggaatgta tctaggtgaa
atcgtaagga gagtgcttct taaaatatct ttgcagtctt 1020caatttttgg caatctagat
cagaccaagc tcaaaacccg ctttattctg aggactcctg 1080atatttccgt gatgcatcat
gatggaacac ctgatctcag aattgtggct gaaaaacttg 1140cagataacct gaagatcaca
gacacatcct tagaaacaag gaagatggtt gtcgaaatct 1200gtgacatcgt cacccgaaga
tcagcccggc tggctgctgc tgggatcgta gggatcctca 1260ggaagatcgg cagaggcgtc
ccaggcgaca agcgaaagtc ggtcatcgcc atcgatggcg 1320gtctatatga acattacacc
gaattccggc agtgcctgga gaccacgctg acggagttgc 1380tcggagaaga ggcgtcgaag
tcggtggctg tcaagcttgc aaacgacggg tctggccttg 1440gagctgccct gattgcagct
gctcattctc agtacctgaa ttgattcccc atagacagag 1500ttccaacttt tattagtgac
tcagtgctgt tgttatagtt tatagaatgt ggtgaagttc 1560tgtatgtttc gaaggagcct
gatggactct acaaacaaaa ttttcagaaa tgaaatcgaa 1620gccaaatgtg ttggtggtgt
tgcaatttgc agtatctact ctgaattatt ataggagaaa 1680atagttctga acttttgttg
taccggataa agtagttctg aactttgttg tcggttattt 1740ttggttcagt acttacatca
ataattcctt catatcacaa aaaaaagttt gtatcaacgg 1800atttaaattt tgaaat
181660463PRTOryza sativa
60Met Val Ala Ala Ala Val Ala Ala Ala Glu Gln Val Val Ala Ala Leu 1
5 10 15 Arg Glu Glu Cys
Ala Thr Pro Ala Ala Arg Leu Asp Gly Val Ala Ala 20
25 30 Ala Met Ala Gly Glu Met Ala Ala Gly
Leu Ala Glu Glu Gly Gly Ser 35 40
45 Lys Ile Lys Met Ile Val Ser Tyr Val Asp Asn Leu Pro Asn
Gly Thr 50 55 60
Glu Glu Gly Leu Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg 65
70 75 80 Val Leu Arg Val Gln
Leu Ala Gly Lys Glu Lys Arg Val Val Lys Arg 85
90 95 Glu Ser Arg Glu Val Ser Ile Pro Pro His
Leu Met Ser Gly Asn Ser 100 105
110 Ser Glu Leu Phe Gly Phe Ile Ala Ser Ala Leu Ala Lys Phe Val
Ala 115 120 125 Asp
Glu Gly His Asn Ala Val Phe Asn Asp Arg Gln Arg Glu Leu Gly 130
135 140 Phe Thr Phe Ser Phe Pro
Val Arg Gln Thr Ser Ile Ala Ser Gly Thr 145 150
155 160 Leu Ile Lys Trp Thr Lys Ala Phe Ser Ile Asp
Asp Ala Val Gly Glu 165 170
175 Asp Val Val Ala Glu Leu Gln Met Ala Met Glu Lys Gln Gly Leu Asp
180 185 190 Met Arg
Val Ser Ala Leu Thr Asn Asp Thr Val Gly Thr Leu Ala Ala 195
200 205 Gly Ser Tyr Tyr Asp Glu Asp
Ile Val Val Gly Val Ile Leu Gly Thr 210 215
220 Gly Ser Asn Ala Ala Tyr Leu Glu Lys Ala Asn Ala
Ile Pro Lys Leu 225 230 235
240 Glu Gly Glu Leu Pro Lys Ser Gly Asn Met Val Ile Asn Thr Glu Trp
245 250 255 Gly Asn Phe
Ser Ser Ser Cys Leu Pro Ile Thr Glu Tyr Asp Glu Ala 260
265 270 Leu Asp Lys Glu Ser Leu Asn Pro
Gly Glu Gln Ile Phe Glu Lys Leu 275 280
285 Ile Ser Gly Met Tyr Leu Gly Glu Ile Val Arg Arg Val
Leu Leu Lys 290 295 300
Ile Ser Leu Gln Ser Ser Ile Phe Gly Asn Leu Asp Gln Thr Lys Leu 305
310 315 320 Lys Thr Arg Phe
Ile Leu Arg Thr Pro Asp Ile Ser Val Met His His 325
330 335 Asp Gly Thr Pro Asp Leu Arg Ile Val
Ala Glu Lys Leu Ala Asp Asn 340 345
350 Leu Lys Ile Thr Asp Thr Ser Leu Glu Thr Arg Lys Met Val
Val Glu 355 360 365
Ile Cys Asp Ile Val Thr Arg Arg Ser Ala Arg Leu Ala Ala Ala Gly 370
375 380 Ile Val Gly Ile Leu
Arg Lys Ile Gly Arg Gly Val Pro Gly Asp Lys 385 390
395 400 Arg Lys Ser Val Ile Ala Ile Asp Gly Gly
Leu Tyr Glu His Tyr Thr 405 410
415 Glu Phe Arg Gln Cys Leu Glu Thr Thr Leu Thr Glu Leu Leu Gly
Glu 420 425 430 Glu
Ala Ser Lys Ser Val Ala Val Lys Leu Ala Asn Asp Gly Ser Gly 435
440 445 Leu Gly Ala Ala Leu Ile
Ala Ala Ala His Ser Gln Tyr Leu Asn 450 455
460 611835DNAOryza sativa 61cactgaaagg gatcaactaa
actactgatg aaaaaaagtt gagtgcttta gacaaaaaca 60caaaactttc acgtgcctcc
ctccaatccc agattatttt tctatagatc gatcgatcga 120gccccgtgaa tgcgattctg
cgcgcatttc cagagcgtct tttgatcgtg gttgggtttg 180tctgaaccga actccaaagg
tctcgccttt cttgctctga agcttttctt tagcttttcg 240attcgcctgt cggggaccgg
gatcatggcg gcggtggagg cggagaaggt ggtggcggag 300ctgcgggaga ggtgcgcgac
gcgggcgtcg ctgctgcggg atgtggcggc ggcgatggcc 360ggcgagatgg gcgccggcct
ggagaaggag ggtgggagca gggtcaagat gctgctctcc 420tacgtcgaca agctccccac
tgggagagag gatggattgt tctatggatt ggacctagga 480ggaacaaact tccgggtgct
caaggttcat cttggtggca gcaagaagca tgtcgtcaac 540tctgaatcca gggaagtcag
catcccacca cacctgatgt cagggacctc ctcggaattg 600tttggtttca ttgctgggga
attaggcaag tttgttgctg aagaggagga gggtactgac 660atgccaaacg gcaagaagaa
agagctagga ttcaccttct cttttcctgt gaggcaacga 720tctgtggcat cgggtaccct
tgtcaagtgg acaaaggcgt tttccattga tgatgcagta 780ggtgaagatg tggtggctga
actacaaacg gctatggtga aacaaggtct ggacatgcat 840gtagctgcat tgattaatga
tgctgttgga acattggctg gagcaagata ctacgatgaa 900gatgttgtcg caggtgtgat
atttggtact ggcacaaatg ctgcatatgt tgagaaggca 960aatgctatac caaaatggga
aggggagttg cccaattcag gggatatggt catcaatatg 1020gaatggggta atttctattc
atcgcatctt ccagttactg aatacgatga agcattagac 1080aaggaaagct taaaccctgg
agagcagata tatgagaagt taacatcagg aatgtattta 1140ggtgaaatcg taagaagagt
gctgcttaaa ctgtccttgc agtctggaat tttcggttct 1200atagataact ccaagctcaa
aacttgtttc catctgcgga ctccgcacat ttctgcaatg 1260caccatgatg aaacacctga
cctaaagata gtggctgaaa aattgcatca aatcctagag 1320attacacata catccttaga
gataaggaaa atggttgtcg aaatatgcga tatcgtggca 1380aggagggcag ctcggctggc
tgctgctggt gttgcaggga tcctcatgaa gcttggaaga 1440aatggcggca tcaacaatca
gcgctcggtc atcgccattg atggaggttt gttcgaacac 1500tacaccaaat tcagggaatg
tttggagagc acgttgggtg agttgctggg agaggaggct 1560tccaagtcgg tagccgtcaa
gcacgcgaac gacgggtcag ggataggggc tgcccttatt 1620gcagcctctc aatctcgatg
aaaatttgct agataaatag tagtatatgc cgtccatttt 1680tgctctggag tgaaaaaaat
tattgaaaaa acctagcttt gttgtatgtg ctgcttgggc 1740atggtggaac tggaagcagt
ctgtaaccaa aatgaggctt gatatactgt cgccctgcat 1800agttgtaacc tgaaataaaa
ttggaactgc aaaac 183562458PRTOryza sativa
62Met Ala Ala Val Glu Ala Glu Lys Val Val Ala Glu Leu Arg Glu Arg 1
5 10 15 Cys Ala Thr Arg
Ala Ser Leu Leu Arg Asp Val Ala Ala Ala Met Ala 20
25 30 Gly Glu Met Gly Ala Gly Leu Glu Lys
Glu Gly Gly Ser Arg Val Lys 35 40
45 Met Leu Leu Ser Tyr Val Asp Lys Leu Pro Thr Gly Arg Glu
Asp Gly 50 55 60
Leu Phe Tyr Gly Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Lys 65
70 75 80 Val His Leu Gly Gly
Ser Lys Lys His Val Val Asn Ser Glu Ser Arg 85
90 95 Glu Val Ser Ile Pro Pro His Leu Met Ser
Gly Thr Ser Ser Glu Leu 100 105
110 Phe Gly Phe Ile Ala Gly Glu Leu Gly Lys Phe Val Ala Glu Glu
Glu 115 120 125 Glu
Gly Thr Asp Met Pro Asn Gly Lys Lys Lys Glu Leu Gly Phe Thr 130
135 140 Phe Ser Phe Pro Val Arg
Gln Arg Ser Val Ala Ser Gly Thr Leu Val 145 150
155 160 Lys Trp Thr Lys Ala Phe Ser Ile Asp Asp Ala
Val Gly Glu Asp Val 165 170
175 Val Ala Glu Leu Gln Thr Ala Met Val Lys Gln Gly Leu Asp Met His
180 185 190 Val Ala
Ala Leu Ile Asn Asp Ala Val Gly Thr Leu Ala Gly Ala Arg 195
200 205 Tyr Tyr Asp Glu Asp Val Val
Ala Gly Val Ile Phe Gly Thr Gly Thr 210 215
220 Asn Ala Ala Tyr Val Glu Lys Ala Asn Ala Ile Pro
Lys Trp Glu Gly 225 230 235
240 Glu Leu Pro Asn Ser Gly Asp Met Val Ile Asn Met Glu Trp Gly Asn
245 250 255 Phe Tyr Ser
Ser His Leu Pro Val Thr Glu Tyr Asp Glu Ala Leu Asp 260
265 270 Lys Glu Ser Leu Asn Pro Gly Glu
Gln Ile Tyr Glu Lys Leu Thr Ser 275 280
285 Gly Met Tyr Leu Gly Glu Ile Val Arg Arg Val Leu Leu
Lys Leu Ser 290 295 300
Leu Gln Ser Gly Ile Phe Gly Ser Ile Asp Asn Ser Lys Leu Lys Thr 305
310 315 320 Cys Phe His Leu
Arg Thr Pro His Ile Ser Ala Met His His Asp Glu 325
330 335 Thr Pro Asp Leu Lys Ile Val Ala Glu
Lys Leu His Gln Ile Leu Glu 340 345
350 Ile Thr His Thr Ser Leu Glu Ile Arg Lys Met Val Val Glu
Ile Cys 355 360 365
Asp Ile Val Ala Arg Arg Ala Ala Arg Leu Ala Ala Ala Gly Val Ala 370
375 380 Gly Ile Leu Met Lys
Leu Gly Arg Asn Gly Gly Ile Asn Asn Gln Arg 385 390
395 400 Ser Val Ile Ala Ile Asp Gly Gly Leu Phe
Glu His Tyr Thr Lys Phe 405 410
415 Arg Glu Cys Leu Glu Ser Thr Leu Gly Glu Leu Leu Gly Glu Glu
Ala 420 425 430 Ser
Lys Ser Val Ala Val Lys His Ala Asn Asp Gly Ser Gly Ile Gly 435
440 445 Ala Ala Leu Ile Ala Ala
Ser Gln Ser Arg 450 455 631844DNAOryza
sativa 63gaactcgcgt tgacctattg actcgtgtcg ccgtctcgcc gagcgaaccc
gacgtcgtcg 60gcgatgagga aggcggcggc gctggcgtcc gcggcgatgg ccgcagcagc
agtagcggta 120gtctccacgg tgttgcacca gaggcaacgt cgggcggcga agcggtcaga
gcgcgcggag 180gccgtgctgc tgcgggacct gcaggagcgg tgcgccgcgc cggtggagct
gctgcggcag 240gtggcggacg cgatggccgc ggagatgcgc gcggggctcg ccgccgaggg
cgggagcgac 300ctccagatgc tcgtcaccta cgttgactcc ctcccctccg ggggtgagaa
agggatgttt 360tatgcacttg accttggagg aacaaatttc cgtgttttac gagttcaatt
aggaggcaaa 420gaacgtcgaa ttatcaagca agactcagaa gggatatcca ttccacaaca
tttaatgtcc 480agcagttcac atgagttgtt tgattttgtt gctgtggctt tagcaaaatt
tgttgcctct 540gaaggtgaag actgccatct tcctgagggt acccaaagag aactaggttt
tacattctcc 600tttccagtga aacaaaaatc attggcatct ggcactctta tcaagtggac
gaagagtttt 660gcaattgatg aaatggtcgg caaggatgtt gtggctgaat taaacatggc
tatcagaagt 720caaggacttg atatgaaagt cacagcattg gttaatgata cagtagggac
attagctgct 780gggagatatg tgaatcatga tactattgct gctgttatac tgggaacagg
tagtaatgca 840gcgtacatag atcatgcaga tgcaattcca aaatggcatg gatccctgcc
caagtctgga 900aatatggtaa taaacatgga atggggtaac tttaagtcct cacatcttcc
acttactgaa 960tttgatcaag agttggatgc agaaagtttg aaccctggca aacaggttta
cgagaaatcg 1020atttctggta tgtatatggg ggaacttgtt cgaagaatct tactaaagat
ggctcaagaa 1080actcgcattt ttggtgataa tatacctcca aaacttgaga gaccatacat
cttaaggaca 1140cttgacatgc tgatcatgca tcatgataca tcatctgatc tcagaacagt
tgccaacaag 1200ttgaaagaag tcttggggat cgaatatacc tctttcacga cgaggaaact
ggttttggat 1260gtttgtgagg ccattgcgac acgcggtgca cggcttgctg ctgctgggat
atatggcatt 1320atccaaaagc ttggtcagca ttctgacagc cccagtacga gaaggtccgt
gattgctgtg 1380gatggagggg tctataaata ctacactttc ttcagccagt gcatggagag
cactctgagt 1440gacatgcttg ggcaggagct ggccccctct gttatgatca agcatgtcaa
tgatggctca 1500ggcgttgggg cagctctcct ggcagcctct tattctcaat accaccaggc
tgaatctgca 1560gatagttcat aatattctaa aaaaaagaag ctgaatctgc agatagctct
taatattctg 1620aaaaaactgt caaaaaataa tattctgaaa aaaaactgtg tattaaggtg
ataaacaata 1680ggttttggag caattttttt tttaagataa tggattaaac cggcctctac
atccaaacga 1740gattctagag caatagcagc tatacagttt gcctaagggc taaatatctt
gtattttgca 1800aatgtcaatt gtacatgaac tctatctgca atatctgttc agtg
184464502PRTOryza sativa 64Met Arg Lys Ala Ala Ala Leu Ala Ser
Ala Ala Met Ala Ala Ala Ala 1 5 10
15 Val Ala Val Val Ser Thr Val Leu His Gln Arg Gln Arg Arg
Ala Ala 20 25 30
Lys Arg Ser Glu Arg Ala Glu Ala Val Leu Leu Arg Asp Leu Gln Glu
35 40 45 Arg Cys Ala Ala
Pro Val Glu Leu Leu Arg Gln Val Ala Asp Ala Met 50
55 60 Ala Ala Glu Met Arg Ala Gly Leu
Ala Ala Glu Gly Gly Ser Asp Leu 65 70
75 80 Gln Met Leu Val Thr Tyr Val Asp Ser Leu Pro Ser
Gly Gly Glu Lys 85 90
95 Gly Met Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val Leu
100 105 110 Arg Val Gln
Leu Gly Gly Lys Glu Arg Arg Ile Ile Lys Gln Asp Ser 115
120 125 Glu Gly Ile Ser Ile Pro Gln His
Leu Met Ser Ser Ser Ser His Glu 130 135
140 Leu Phe Asp Phe Val Ala Val Ala Leu Ala Lys Phe Val
Ala Ser Glu 145 150 155
160 Gly Glu Asp Cys His Leu Pro Glu Gly Thr Gln Arg Glu Leu Gly Phe
165 170 175 Thr Phe Ser Phe
Pro Val Lys Gln Lys Ser Leu Ala Ser Gly Thr Leu 180
185 190 Ile Lys Trp Thr Lys Ser Phe Ala Ile
Asp Glu Met Val Gly Lys Asp 195 200
205 Val Val Ala Glu Leu Asn Met Ala Ile Arg Ser Gln Gly Leu
Asp Met 210 215 220
Lys Val Thr Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala Ala Gly 225
230 235 240 Arg Tyr Val Asn His
Asp Thr Ile Ala Ala Val Ile Leu Gly Thr Gly 245
250 255 Ser Asn Ala Ala Tyr Ile Asp His Ala Asp
Ala Ile Pro Lys Trp His 260 265
270 Gly Ser Leu Pro Lys Ser Gly Asn Met Val Ile Asn Met Glu Trp
Gly 275 280 285 Asn
Phe Lys Ser Ser His Leu Pro Leu Thr Glu Phe Asp Gln Glu Leu 290
295 300 Asp Ala Glu Ser Leu Asn
Pro Gly Lys Gln Val Tyr Glu Lys Ser Ile 305 310
315 320 Ser Gly Met Tyr Met Gly Glu Leu Val Arg Arg
Ile Leu Leu Lys Met 325 330
335 Ala Gln Glu Thr Arg Ile Phe Gly Asp Asn Ile Pro Pro Lys Leu Glu
340 345 350 Arg Pro
Tyr Ile Leu Arg Thr Leu Asp Met Leu Ile Met His His Asp 355
360 365 Thr Ser Ser Asp Leu Arg Thr
Val Ala Asn Lys Leu Lys Glu Val Leu 370 375
380 Gly Ile Glu Tyr Thr Ser Phe Thr Thr Arg Lys Leu
Val Leu Asp Val 385 390 395
400 Cys Glu Ala Ile Ala Thr Arg Gly Ala Arg Leu Ala Ala Ala Gly Ile
405 410 415 Tyr Gly Ile
Ile Gln Lys Leu Gly Gln His Ser Asp Ser Pro Ser Thr 420
425 430 Arg Arg Ser Val Ile Ala Val Asp
Gly Gly Val Tyr Lys Tyr Tyr Thr 435 440
445 Phe Phe Ser Gln Cys Met Glu Ser Thr Leu Ser Asp Met
Leu Gly Gln 450 455 460
Glu Leu Ala Pro Ser Val Met Ile Lys His Val Asn Asp Gly Ser Gly 465
470 475 480 Val Gly Ala Ala
Leu Leu Ala Ala Ser Tyr Ser Gln Tyr His Gln Ala 485
490 495 Glu Ser Ala Asp Ser Ser
500 651585DNAOryza sativa 65gtggagtgat cgatcgactc gggcgcgggg
agggggaggg gaggggaggt gggggggaat 60ggaggggagg gcggcggggt gggtgagggt
ggcggcggtg gggtgggcgg tggcggcgtg 120cgcggtggcg gcggggatgg tggcgaggcg
aggggcggcg agggtgcggt ggaacagggc 180ggtggcggtg gtgcgggacc tggaggagcg
gtgcgccacg cccgcggagc tgctgcagcg 240ggtggtcaac tcgctggcca tcgagatgtt
cgccggcctc gcctccgacg gcgggagcaa 300ggtgcggatg ctgctcacct gcgtcgacgc
gctccccgac gggagcgagg aaggcatcag 360ttatgccatt gatcttggag gaacaagctt
tagagtcctg aaagtagaac ttggcgcagg 420gtctacaatc atcaatcgaa aagttgaaca
tcagcctatc cccgaaaatt tgactaaggg 480tacaagcgat gatttgttca atttcattgc
ctcggcactg aagaatttta ttgaaagaga 540gggtggggag gttgagggga gggcacttgg
ttttacattt tctttccctg tgagacagac 600ttccatttcc tcggggacat taattcgatg
gactaaagaa ttttcaattg aagaggctgt 660cgggaaagat gttgctcagt gcctaaatga
agcccttgct aggaatggac tcaatatgaa 720ggtcaatgta ttggtgaaca atactgtggg
gacattagct ctcgggcatt attatgatga 780tgacacagta gctgctgtga ttattggagc
tggcaccaat gcttgctata ttgaacgcaa 840cgatgcaatt attaaatctt tgggtcgcgt
taccaattct gaacgaacgg tagtaaatgt 900ggaatggggt agttttcggc ctccacaaat
agaattgact ccttatgata tatgcttcaa 960caacgaaaca tggaattatt atgaccaggg
ttttgagaaa atgatctctg gtgtgtatct 1020tggggaaatt gcaagattgg tgttccaaaa
aatggctgaa gagtcagata tatttggtac 1080tgctgttgat ggtctatcga cccctttcgt
cttaagtaca ccaaacttag ctgctatccg 1140cgaggatgat tccccggact tgagagaagt
cggcaagata cttgaggaac atcttaagct 1200accagatgtt cccctcaaga cccggaagct
tgttgccaga gtctccgaca tcatcacccg 1260gagagctgcc cgcctagcag cggctgcgat
tgtcgcgata ctgcaaaaga tcggctgcga 1320tggaaccctt tgtggttcaa ctcaggttcg
aacaatgcga ggcgtgcgaa gaagaacggt 1380ggtcgcgatc gagggcggcc tcttcgaagg
ctactcggtc ttcagagagt atctgaatga 1440agctctagtt gagatccttg gagaggagat
tgcagccact gttagtctta gggtgatgga 1500ggagggatct gggactggtg ctgccctcct
tgcagctgca tattcatcgg ctaggcagaa 1560gaactccgag taggcatgag acaca
158566504PRTOryza sativa 66Met Glu Gly
Arg Ala Ala Gly Trp Val Arg Val Ala Ala Val Gly Trp 1 5
10 15 Ala Val Ala Ala Cys Ala Val Ala
Ala Gly Met Val Ala Arg Arg Gly 20 25
30 Ala Ala Arg Val Arg Trp Asn Arg Ala Val Ala Val Val
Arg Asp Leu 35 40 45
Glu Glu Arg Cys Ala Thr Pro Ala Glu Leu Leu Gln Arg Val Val Asn 50
55 60 Ser Leu Ala Ile
Glu Met Phe Ala Gly Leu Ala Ser Asp Gly Gly Ser 65 70
75 80 Lys Val Arg Met Leu Leu Thr Cys Val
Asp Ala Leu Pro Asp Gly Ser 85 90
95 Glu Glu Gly Ile Ser Tyr Ala Ile Asp Leu Gly Gly Thr Ser
Phe Arg 100 105 110
Val Leu Lys Val Glu Leu Gly Ala Gly Ser Thr Ile Ile Asn Arg Lys
115 120 125 Val Glu His Gln
Pro Ile Pro Glu Asn Leu Thr Lys Gly Thr Ser Asp 130
135 140 Asp Leu Phe Asn Phe Ile Ala Ser
Ala Leu Lys Asn Phe Ile Glu Arg 145 150
155 160 Glu Gly Gly Glu Val Glu Gly Arg Ala Leu Gly Phe
Thr Phe Ser Phe 165 170
175 Pro Val Arg Gln Thr Ser Ile Ser Ser Gly Thr Leu Ile Arg Trp Thr
180 185 190 Lys Glu Phe
Ser Ile Glu Glu Ala Val Gly Lys Asp Val Ala Gln Cys 195
200 205 Leu Asn Glu Ala Leu Ala Arg Asn
Gly Leu Asn Met Lys Val Asn Val 210 215
220 Leu Val Asn Asn Thr Val Gly Thr Leu Ala Leu Gly His
Tyr Tyr Asp 225 230 235
240 Asp Asp Thr Val Ala Ala Val Ile Ile Gly Ala Gly Thr Asn Ala Cys
245 250 255 Tyr Ile Glu Arg
Asn Asp Ala Ile Ile Lys Ser Leu Gly Arg Val Thr 260
265 270 Asn Ser Glu Arg Thr Val Val Asn Val
Glu Trp Gly Ser Phe Arg Pro 275 280
285 Pro Gln Ile Glu Leu Thr Pro Tyr Asp Ile Cys Phe Asn Asn
Glu Thr 290 295 300
Trp Asn Tyr Tyr Asp Gln Gly Phe Glu Lys Met Ile Ser Gly Val Tyr 305
310 315 320 Leu Gly Glu Ile Ala
Arg Leu Val Phe Gln Lys Met Ala Glu Glu Ser 325
330 335 Asp Ile Phe Gly Thr Ala Val Asp Gly Leu
Ser Thr Pro Phe Val Leu 340 345
350 Ser Thr Pro Asn Leu Ala Ala Ile Arg Glu Asp Asp Ser Pro Asp
Leu 355 360 365 Arg
Glu Val Gly Lys Ile Leu Glu Glu His Leu Lys Leu Pro Asp Val 370
375 380 Pro Leu Lys Thr Arg Lys
Leu Val Ala Arg Val Ser Asp Ile Ile Thr 385 390
395 400 Arg Arg Ala Ala Arg Leu Ala Ala Ala Ala Ile
Val Ala Ile Leu Gln 405 410
415 Lys Ile Gly Cys Asp Gly Thr Leu Cys Gly Ser Thr Gln Val Arg Thr
420 425 430 Met Arg
Gly Val Arg Arg Arg Thr Val Val Ala Ile Glu Gly Gly Leu 435
440 445 Phe Glu Gly Tyr Ser Val Phe
Arg Glu Tyr Leu Asn Glu Ala Leu Val 450 455
460 Glu Ile Leu Gly Glu Glu Ile Ala Ala Thr Val Ser
Leu Arg Val Met 465 470 475
480 Glu Glu Gly Ser Gly Thr Gly Ala Ala Leu Leu Ala Ala Ala Tyr Ser
485 490 495 Ser Ala Arg
Gln Lys Asn Ser Glu 500 671494DNASorghum
bicolor 67atggggcggg tcgggctcgg cgtggcggcg ggctgcgcgg ccgccacgtg
cgcgatcgcc 60gccgcgctgg tggcgcgcag ggcgtcggcg cgggcgcgct ggcgccgcgc
cgtcgcgctg 120ctcagggagt tcgaggaagg ctgcgccacg ccgacgccgc gcctgcgcca
ggtcgtcgac 180gccatggtcg tcgagatgca cgcgggcctc gcctccgacg gcggcagcaa
gctcaagatg 240ctgctcacct tcgtcgacgc gctccccgcc ggaaatgaac aaggcacata
ttattccatc 300gatcttggag gaacaaactt tagagtgttg agagttgaag ttggtgctgt
gtctgtggtg 360accagtcggg aggttaaact tcccatccct gaggaattga ccaagggtac
aattgaggag 420ctattcaact ttgttgccat gaccctaaag gaatttgtag agacagaaga
tgttaaagat 480gaacaaaggg cgcttggttt cacattttct ttcccagtta gacaaacttc
agtgtcttca 540gggtcattaa ttaggtggac taaaggtttt ttgattgaag atgcggttgg
gaaagatgtg 600gctcaatgct taaatgaagc tcttgctagg agtggactaa atgtgcgagt
tactgcactg 660gtgaacgaca ccgtggggac attagctcta ggacattatt atgatgagga
tacagtggct 720gctgtgatca tcggtgctgg caccaatgct tgctatattg aacgcactga
tgcaattatt 780aaatgtcagg gtcttcttac aaactctggt ggcatggtag taaacatgga
atggggcaat 840ttctggtcat cacatttgcc aagaactcct tatgacatct cccttgatga
tgagacgcaa 900aatcgcaatg atcaggggtt tgagaaaatg atctctggga tttatctcgg
ggaaattgca 960aggctggtgc tgcatcgcat ggctctagaa tcagatgttt ttggcgatgc
tgctgatcat 1020ctatctaccc ccttcacatt gagcacacca cttctggctg caattcgcaa
ggacgattca 1080ccagatctga gtgaagtcag aaggatactg caagaacatc tgaagataat
ggacactccc 1140ctgaaaactc gaaggctagt cgtcaaagtc tgcgacattg tcacccgaag
agctgcacgc 1200ctagctgctg ctggtattgt cgggatactg aaaaagctcg gtcgggatgg
gagcggcgtg 1260gcttcaagcg ggagaacacg agggcagctg aggcggacgg tggttgcgat
cgagggtggc 1320ctgtatgagg gctacccagt gttcagggag tacctagatg aagctctggt
ggagatcttg 1380ggggaggagg tggcgcagac ggtggcgcta agggtgacag aggacgggtc
tggggctggc 1440gctgccctcc tcgccgccgt acattcgtcg aatagacagc aaggttccat
atag 149468497PRTSorghum bicolor 68Met Gly Arg Val Gly Leu Gly
Val Ala Ala Gly Cys Ala Ala Ala Thr 1 5
10 15 Cys Ala Ile Ala Ala Ala Leu Val Ala Arg Arg
Ala Ser Ala Arg Ala 20 25
30 Arg Trp Arg Arg Ala Val Ala Leu Leu Arg Glu Phe Glu Glu Gly
Cys 35 40 45 Ala
Thr Pro Thr Pro Arg Leu Arg Gln Val Val Asp Ala Met Val Val 50
55 60 Glu Met His Ala Gly Leu
Ala Ser Asp Gly Gly Ser Lys Leu Lys Met 65 70
75 80 Leu Leu Thr Phe Val Asp Ala Leu Pro Ala Gly
Asn Glu Gln Gly Thr 85 90
95 Tyr Tyr Ser Ile Asp Leu Gly Gly Thr Asn Phe Arg Val Leu Arg Val
100 105 110 Glu Val
Gly Ala Val Ser Val Val Thr Ser Arg Glu Val Lys Leu Pro 115
120 125 Ile Pro Glu Glu Leu Thr Lys
Gly Thr Ile Glu Glu Leu Phe Asn Phe 130 135
140 Val Ala Met Thr Leu Lys Glu Phe Val Glu Thr Glu
Asp Val Lys Asp 145 150 155
160 Glu Gln Arg Ala Leu Gly Phe Thr Phe Ser Phe Pro Val Arg Gln Thr
165 170 175 Ser Val Ser
Ser Gly Ser Leu Ile Arg Trp Thr Lys Gly Phe Leu Ile 180
185 190 Glu Asp Ala Val Gly Lys Asp Val
Ala Gln Cys Leu Asn Glu Ala Leu 195 200
205 Ala Arg Ser Gly Leu Asn Val Arg Val Thr Ala Leu Val
Asn Asp Thr 210 215 220
Val Gly Thr Leu Ala Leu Gly His Tyr Tyr Asp Glu Asp Thr Val Ala 225
230 235 240 Ala Val Ile Ile
Gly Ala Gly Thr Asn Ala Cys Tyr Ile Glu Arg Thr 245
250 255 Asp Ala Ile Ile Lys Cys Gln Gly Leu
Leu Thr Asn Ser Gly Gly Met 260 265
270 Val Val Asn Met Glu Trp Gly Asn Phe Trp Ser Ser His Leu
Pro Arg 275 280 285
Thr Pro Tyr Asp Ile Ser Leu Asp Asp Glu Thr Gln Asn Arg Asn Asp 290
295 300 Gln Gly Phe Glu Lys
Met Ile Ser Gly Ile Tyr Leu Gly Glu Ile Ala 305 310
315 320 Arg Leu Val Leu His Arg Met Ala Leu Glu
Ser Asp Val Phe Gly Asp 325 330
335 Ala Ala Asp His Leu Ser Thr Pro Phe Thr Leu Ser Thr Pro Leu
Leu 340 345 350 Ala
Ala Ile Arg Lys Asp Asp Ser Pro Asp Leu Ser Glu Val Arg Arg 355
360 365 Ile Leu Gln Glu His Leu
Lys Ile Met Asp Thr Pro Leu Lys Thr Arg 370 375
380 Arg Leu Val Val Lys Val Cys Asp Ile Val Thr
Arg Arg Ala Ala Arg 385 390 395
400 Leu Ala Ala Ala Gly Ile Val Gly Ile Leu Lys Lys Leu Gly Arg Asp
405 410 415 Gly Ser
Gly Val Ala Ser Ser Gly Arg Thr Arg Gly Gln Leu Arg Arg 420
425 430 Thr Val Val Ala Ile Glu Gly
Gly Leu Tyr Glu Gly Tyr Pro Val Phe 435 440
445 Arg Glu Tyr Leu Asp Glu Ala Leu Val Glu Ile Leu
Gly Glu Glu Val 450 455 460
Ala Gln Thr Val Ala Leu Arg Val Thr Glu Asp Gly Ser Gly Ala Gly 465
470 475 480 Ala Ala Leu
Leu Ala Ala Val His Ser Ser Asn Arg Gln Gln Gly Ser 485
490 495 Ile 692045DNASorghum bicolor
69cttggtgcta gcctcctttt cccttccccc ttgaatcgaa ctgttgggag ctcttcgctg
60tcctaccatt cccccctcgc tcttttttag cattttgctt gggctctcga ctcctccgaa
120aatccagcct tttgtttgct tttctgtctc ctgctgattg gtgtccccgg atccaggatt
180cttgatttgc tcctcggggg ttggggccat ggcgactgct gcgctggcaa tggcagagca
240ggtggtggcc gacctccgag cgaagtgtga ggcgccgccg ccgatgctgc gcgaggtggc
300ggcggagatg gcccgcgaga tgggcgcggg actggagaag gaaggcggga gcagggtcaa
360gatgctcctc tcctacgttg ataagctccc cacaggggga gaagagggat tattctatgg
420attggaccta ggagggacga acttccgcgt cttgaaagtg gaactgggtg ggaatgagaa
480gcacgtcgtg gaccgtgact ccagagaagt cattattcct ccacatttga tgtcagggag
540ctcctcggag ctttttggtt tcattgcttc tgaattggcc aagtttgttg ttgatgatga
600gaagttcatc aatgttttga atggaaagaa gcgagaacta gggttcacat tttcattccc
660agtgaagcag cgttctgttg cttccggtac gcttgtcaag tggacaaagg cattttccat
720taatgatgct gttggtgaag atgtggtggc taaactgcaa acagctatgg agaagcaagg
780tctggacatg catgtagctg cattgattaa tgatgctgtt gggactctgg ctggagcaag
840gtactacgac aaagatgttg tcgctggtgt aatatttggc actggcacaa atgcagcata
900tgttgagaag gcaaatgcta ttccaaaatg ggagggtgag ctgcccaatt caggagatat
960ggtcatcaac atggaatggg gtaacttctg ctcagcctat ctcccaatca ctgaatatga
1020ccaagaatta gataaggaga gcttaaatcc aggagaacag atttatgaga agttaacatc
1080agggatgtat ttgggcgaaa ttgtaaggag ggtgctcctt aaaatatcat tgcagtctgc
1140gatttttggc aatattgacc acactaagct cgaaaccccg ttccttctgc ggactccaca
1200tatttctgca atgcaccatg atgaaacacc tgatctgaag attgtggcaa aaaaactgga
1260agaaaaccta gagattacag gcgcatcctt agaggctcga aaattggtgg ttgaaatttg
1320tgacattgtg gcaacaagag ctgcccggct ggccgctgca gggcttgcag ggatcctcat
1380gaagctcggg agagattgca gtgtcgagga tcaacgatca gtcatcgcca tcgatggagg
1440attgttcgag cactacacca aattccgcca atgcttggag accacactgg gcgagctgct
1500aggagatgag gcgtctaagg cggtggccgt caagcatgca gatgatggtt caggaatagg
1560tgctgccctg attgcagctt cacaatctct gtacaaaaat gacttagtgg ccgtcaagca
1620tgcagatgac aagcatgcag atgacaagca tgaagatgca gatgacaagc atgaagatga
1680cggtaaagga gtcaagcatg cagatgacgg ttcagaaata ggtgctgccc tgattgcagc
1740ctcgcaatct cagtagagaa atgtcctcga aatctcagta gagaaatttc gagtgatata
1800gtagtatcag ccatgggtgc ttaccataga tgtaggaaag actagctagc aagtagctat
1860aatgtctttc caacatctta acagtccggc atctgcagtt agcatgctgg acctggatgt
1920aatctataac caaaacgagg cttggcagct ggatgcaatc tgtaaccaaa acgaggcttg
1980ggaagtgctg ttcatagttc caaacttcca ataaaagcgg agttgcagat gtctcctaat
2040tcgac
204570515PRTSorghum bicolor 70Met Ala Thr Ala Ala Leu Ala Met Ala Glu Gln
Val Val Ala Asp Leu 1 5 10
15 Arg Ala Lys Cys Glu Ala Pro Pro Pro Met Leu Arg Glu Val Ala Ala
20 25 30 Glu Met
Ala Arg Glu Met Gly Ala Gly Leu Glu Lys Glu Gly Gly Ser 35
40 45 Arg Val Lys Met Leu Leu Ser
Tyr Val Asp Lys Leu Pro Thr Gly Gly 50 55
60 Glu Glu Gly Leu Phe Tyr Gly Leu Asp Leu Gly Gly
Thr Asn Phe Arg 65 70 75
80 Val Leu Lys Val Glu Leu Gly Gly Asn Glu Lys His Val Val Asp Arg
85 90 95 Asp Ser Arg
Glu Val Ile Ile Pro Pro His Leu Met Ser Gly Ser Ser 100
105 110 Ser Glu Leu Phe Gly Phe Ile Ala
Ser Glu Leu Ala Lys Phe Val Val 115 120
125 Asp Asp Glu Lys Phe Ile Asn Val Leu Asn Gly Lys Lys
Arg Glu Leu 130 135 140
Gly Phe Thr Phe Ser Phe Pro Val Lys Gln Arg Ser Val Ala Ser Gly 145
150 155 160 Thr Leu Val Lys
Trp Thr Lys Ala Phe Ser Ile Asn Asp Ala Val Gly 165
170 175 Glu Asp Val Val Ala Lys Leu Gln Thr
Ala Met Glu Lys Gln Gly Leu 180 185
190 Asp Met His Val Ala Ala Leu Ile Asn Asp Ala Val Gly Thr
Leu Ala 195 200 205
Gly Ala Arg Tyr Tyr Asp Lys Asp Val Val Ala Gly Val Ile Phe Gly 210
215 220 Thr Gly Thr Asn Ala
Ala Tyr Val Glu Lys Ala Asn Ala Ile Pro Lys 225 230
235 240 Trp Glu Gly Glu Leu Pro Asn Ser Gly Asp
Met Val Ile Asn Met Glu 245 250
255 Trp Gly Asn Phe Cys Ser Ala Tyr Leu Pro Ile Thr Glu Tyr Asp
Gln 260 265 270 Glu
Leu Asp Lys Glu Ser Leu Asn Pro Gly Glu Gln Ile Tyr Glu Lys 275
280 285 Leu Thr Ser Gly Met Tyr
Leu Gly Glu Ile Val Arg Arg Val Leu Leu 290 295
300 Lys Ile Ser Leu Gln Ser Ala Ile Phe Gly Asn
Ile Asp His Thr Lys 305 310 315
320 Leu Glu Thr Pro Phe Leu Leu Arg Thr Pro His Ile Ser Ala Met His
325 330 335 His Asp
Glu Thr Pro Asp Leu Lys Ile Val Ala Lys Lys Leu Glu Glu 340
345 350 Asn Leu Glu Ile Thr Gly Ala
Ser Leu Glu Ala Arg Lys Leu Val Val 355 360
365 Glu Ile Cys Asp Ile Val Ala Thr Arg Ala Ala Arg
Leu Ala Ala Ala 370 375 380
Gly Leu Ala Gly Ile Leu Met Lys Leu Gly Arg Asp Cys Ser Val Glu 385
390 395 400 Asp Gln Arg
Ser Val Ile Ala Ile Asp Gly Gly Leu Phe Glu His Tyr 405
410 415 Thr Lys Phe Arg Gln Cys Leu Glu
Thr Thr Leu Gly Glu Leu Leu Gly 420 425
430 Asp Glu Ala Ser Lys Ala Val Ala Val Lys His Ala Asp
Asp Gly Ser 435 440 445
Gly Ile Gly Ala Ala Leu Ile Ala Ala Ser Gln Ser Leu Tyr Lys Asn 450
455 460 Asp Leu Val Ala
Val Lys His Ala Asp Asp Lys His Ala Asp Asp Lys 465 470
475 480 His Glu Asp Ala Asp Asp Lys His Glu
Asp Asp Gly Lys Gly Val Lys 485 490
495 His Ala Asp Asp Gly Ser Glu Ile Gly Ala Ala Leu Ile Ala
Ala Ser 500 505 510
Gln Ser Gln 515 71581DNATriticum aestivum 71gctcaagatg ctcgttagct
acgtcgacaa tctgcccacc ggggatgagc atgggctgtt 60ttatgcactg gatcttggtg
ggaccaactt tcgtgttcta cgggttcagc ttggaggaaa 120ggagaaacgt gctgtccaac
aatatgaaga ggtgcccatt ccacctcatc tgatggttgg 180gacttccacc gaactatttg
atttcattgc ggctgagcta gaaagatttg ttgagactga 240aggagacgat ttccacttgc
ctgagggcag gcatagggaa ctgggtttca ccttttcttt 300cccagtacac caaacatcga
tatcgtcagg caccctcgtt aagtggacaa aaggattttg 360catcaatggc acggttgggg
aagatgtggt ggctgaattg agcagtgcta tggagaggca 420ggggcttgat atgaaagtta
cagctttggt taatgatact gtgggcacgt tggctggtgg 480gatatatgct gataatgatg
tcgtcgctgc tgtaatattg ggcactggga caaatgcagc 540gtatgttgag catgctaata
caattcccaa atggcatggt c 58172193PRTTriticum
aestivum 72Leu Lys Met Leu Val Ser Tyr Val Asp Asn Leu Pro Thr Gly Asp
Glu 1 5 10 15 His
Gly Leu Phe Tyr Ala Leu Asp Leu Gly Gly Thr Asn Phe Arg Val
20 25 30 Leu Arg Val Gln Leu
Gly Gly Lys Glu Lys Arg Ala Val Gln Gln Tyr 35
40 45 Glu Glu Val Pro Ile Pro Pro His Leu
Met Val Gly Thr Ser Thr Glu 50 55
60 Leu Phe Asp Phe Ile Ala Ala Glu Leu Glu Arg Phe Val
Glu Thr Glu 65 70 75
80 Gly Asp Asp Phe His Leu Pro Glu Gly Arg His Arg Glu Leu Gly Phe
85 90 95 Thr Phe Ser Phe
Pro Val His Gln Thr Ser Ile Ser Ser Gly Thr Leu 100
105 110 Val Lys Trp Thr Lys Gly Phe Cys Ile
Asn Gly Thr Val Gly Glu Asp 115 120
125 Val Val Ala Glu Leu Ser Ser Ala Met Glu Arg Gln Gly Leu
Asp Met 130 135 140
Lys Val Thr Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala Gly Gly 145
150 155 160 Ile Tyr Ala Asp Asn
Asp Val Val Ala Ala Val Ile Leu Gly Thr Gly 165
170 175 Thr Asn Ala Ala Tyr Val Glu His Ala Asn
Thr Ile Pro Lys Trp His 180 185
190 Gly 733743DNAPhyscomitrella patens 73gctctctgag cctcacctcc
ctcgctccct tgtcttgctc tgcgatctct cgtcaagctt 60ttttttcccc cctttctttc
tttttctccc gtcctctgtt gtgcttgcgc cagccgatcg 120cctacaccta gcagccgcgg
ttggagtgct gtgtttttag ctttttggag tcctcagatc 180tggcagacag tttgatgtag
ggacgttggt gtaggtttat tcagctgatc gattgagagt 240gcaaaaggcg aagctgtgtg
cgtgtgcgtg tgcgtgtttg ctcgtgagtg tttgggagta 300ttgctaaagg gtagcaatgg
cgatcgggaa ggtactgggg tgtgccgggt ttcagcacag 360tgcagtgcca acgctccgtg
aaccagtgcg gctgagggcg caatgcagac gccggggaaa 420gaccgtgtct atgtcggttc
agaagacgtc gaaaacggta cagcaggcgg agaagatgtc 480gcaggagttc cgtcagtctt
cctctactcc cctgcctctt ttgcaccagg tagcggacgc 540tctcgtgcaa gaaatgtatg
ctggattggc gtcggaaggt ggtagtgatc agctgaagat 600gcttccgacc tatgtcgaaa
acttgccttc tgggtgcgct gcttctcaat accctctcct 660cgtcgatcct tctttcgttg
atgcgaagta gattcttctc ccagacctca gcaccctttg 720tttgtcgaaa gtgcggcatg
caatgctccc tcagtctgtc acggtgtccc gagcaacttc 780taaaaaaaca catattcact
tgttttgcga cgtcaccgcg tagagtgtag cagtcagata 840tttcaatgac ctggtcgtgg
gcgttctctc tttcctgttg ttaaactctc cctatcataa 900catacggcgc agtttgattg
agcttgaaac actgcttgag cacgatcaac gaacattgta 960ctaagagaac attcgcgatc
gctggggaca cgaggccgcg tcttgagaga gatgcctatt 1020gtgaattttg gcattcttac
gaatcccttc gagttttcaa ttgcaggagc gagaaggggc 1080tgttctatgc cgtggatttg
ggcggcacaa acttccgtgt tctgcgagtg gaattgggag 1140gcaaaacagg ccaaattttg
agccaagagt tcaaggaagt ggtaatccct ccagagctta 1200tggtgggcac tggtaaggta
agacttcata cactcaacac ccatttctaa taactcttcg 1260tatgaagagc gggacgagtg
gcgaatgatt ctctgggcat gatgagaagg gccatgattt 1320gattcttatg gcgggatgcc
gtgcaacagg atcttttcga cttcattgct ggtacactcg 1380catcatttgt cgacactgaa
gacgagtcaa ttaaagctca ctttgttcaa tcgggaaaaa 1440ccagggagtc cggtttcgct
ttctccttcc ccgttcgaca gacctcagtc aaatctggca 1500ttgtcatcca ctggacgaag
ggcttcaaag ttgatgatgc ggtgcgccac ttgtgcaggc 1560cttttcactc acaaattcac
tactcataca cacacattta tcccatgaga ggatgcattt 1620gccaatttaa atagcgtttt
cgatacgagt aagcttagat ttgcatcttc tttcttccct 1680tgcggagggg cacttttctg
aatgtttcag ggacttgagg attatccttt acgaacccct 1740gttgtatgcg ctgatggtat
ttgcaaccca aatttagttc aaacttcctt aatggttcat 1800ctttctggca ggtgggcaaa
gacatcgtga agcaatttca ggatgcaatc agcagaagta 1860accatcagat tatgatctca
gccctggtat atctcacttt tcttgaactg cacactctgt 1920ttggatttaa gttctttacg
taatagcgtc catcttctgt ataacctgtg tttgcaatga 1980tagactgaag gatttcttaa
ttttgaatct tttacaggta aacgacaccg ttggcacgtt 2040ggccggaggc agattcaact
tcgacgagga gaccatgatt ggttgcatta ttggaacagg 2100gacaaacgca tgctatgtcg
aacgtgccga tgctgtacat aagtgggacg agccactgcc 2160caaatcagga gaaatggtaa
tcgatttcgc cgcatttctc ggtttgtagg cacctgagca 2220ggaaatcatg ctaacaaacg
tcttcaaatg ggtgatttgc tgacgatcat tgagtatatg 2280agcagttttt tccttacatt
ttgttattac gtgtggcagg ttatcaatat ggaatgggga 2340aatttccgtt caccctacct
gccacgaaca tttgctgatg agactgttga caaagacagc 2400gtcaatccag gggaccaggt
gcgaagtcta ccgagagaac tgagtgcatg aggtgttgaa 2460ttaggtaatt ttcaagagca
gacctattca ccctctttgg tggttttgtg cagtggttcg 2520agaaaatgat ctctggaatg
tacctcggcg agatcgtgcg ccttgttctg gctaggatgg 2580ccaaagaagc ggaactgttt
ggtggcaatg tccccgtgaa gctcttggag cggctcaccc 2640tagggtaatt ccacggtact
tgcagccatt tctcaatgaa ttaatcctca gtaagcctaa 2700tcgtattcgc tcttggcaaa
gcacagatca ttgatccttc actgtttgta cagcacccca 2760catgtttcca aaatacatct
ggacaattca cctgacctcg acgtggttgc caaagttctc 2820aaggacgtct tcgaggtgag
tgcctatgag caaatgcacg ttcgatgaca ttgaatggaa 2880tccgaacaat attgacacca
cttgcactgg tactaagtac ttgattcatg gccccctttt 2940gcagatcgaa accaccacac
ttgaggaaag aaagatagta cacgaggtgt gtgacatcat 3000gggcgagcga ggtggaaggt
tggctgcagc aggtctctac ggaatcctga agaagatcgg 3060aagaaccggc aaatctcgaa
acggatccaa gaagaagacc gtgattgcaa tggacggtgg 3120cctgttcgaa caccacgtcc
gttaccggtc atacatggag gaggcgctcc aggagttgat 3180gggttctgac gcagcgtacg
aggtggcgtt gagactacag aacgatggtt ctggaatcgg 3240tgccgctttg cttgccgctt
cccactcaca cttcaaatag gataaaacgc acacaaagct 3300tagggggatc atggcaggtg
atgagcatcg cctgttttgc atgcctgtac aatattccga 3360gtcctgatga tctgtctatc
ttctgagatg cacccatcgt ctgtaccatt tgtaccggct 3420tatcgtaccg caggaagacg
ccgttcttca agtcgttgtc tatctggtga ccaaaaccgc 3480agagtttcaa gctgagaggc
ctagtagggg tagtgtaggc agagtcaagt tttattgtct 3540ctggtcagtt acgtagaaca
gtttgcggtt gagtctaccc atctcatcta gatctgcact 3600gcacaccttt caataagctg
gtttcacttg aggttgccgt gcgcaagcta gtatctggca 3660cagagatgag aggttcattg
ctgcgcattc tcttagcaat cctcacaatt tccgaggtga 3720ttgtggggtg atggagctgg
gcg 374374513PRTPhyscomitrella
patens 74Met Ala Ile Gly Lys Val Leu Gly Cys Ala Gly Phe Gln His Ser Ala
1 5 10 15 Val Pro
Thr Leu Arg Glu Pro Val Arg Leu Arg Ala Gln Cys Arg Arg 20
25 30 Arg Gly Lys Thr Val Ser Met
Ser Val Gln Lys Thr Ser Lys Thr Val 35 40
45 Gln Gln Ala Glu Lys Met Ser Gln Glu Phe Arg Gln
Ser Ser Ser Thr 50 55 60
Pro Leu Pro Leu Leu His Gln Val Ala Asp Ala Leu Val Gln Glu Met 65
70 75 80 Tyr Ala Gly
Leu Ala Ser Glu Gly Gly Ser Asp Gln Leu Lys Met Leu 85
90 95 Pro Thr Tyr Val Glu Asn Leu Pro
Ser Gly Ser Glu Lys Gly Leu Phe 100 105
110 Tyr Ala Val Asp Leu Gly Gly Thr Asn Phe Arg Val Leu
Arg Val Glu 115 120 125
Leu Gly Gly Lys Thr Gly Gln Ile Leu Ser Gln Glu Phe Lys Glu Val 130
135 140 Val Ile Pro Pro
Glu Leu Met Val Gly Thr Gly Lys Asp Leu Phe Asp 145 150
155 160 Phe Ile Ala Gly Thr Leu Ala Ser Phe
Val Asp Thr Glu Asp Glu Ser 165 170
175 Ile Lys Ala His Phe Val Gln Ser Gly Lys Thr Arg Glu Ser
Gly Phe 180 185 190
Ala Phe Ser Phe Pro Val Arg Gln Thr Ser Val Lys Ser Gly Ile Val
195 200 205 Ile His Trp Thr
Lys Gly Phe Lys Val Asp Asp Ala Val Gly Lys Asp 210
215 220 Ile Val Lys Gln Phe Gln Asp Ala
Ile Ser Arg Ser Asn His Gln Ile 225 230
235 240 Met Ile Ser Ala Leu Val Asn Asp Thr Val Gly Thr
Leu Ala Gly Gly 245 250
255 Arg Phe Asn Phe Asp Glu Glu Thr Met Ile Gly Cys Ile Ile Gly Thr
260 265 270 Gly Thr Asn
Ala Cys Tyr Val Glu Arg Ala Asp Ala Val His Lys Trp 275
280 285 Asp Glu Pro Leu Pro Lys Ser Gly
Glu Met Val Ile Asn Met Glu Trp 290 295
300 Gly Asn Phe Arg Ser Pro Tyr Leu Pro Arg Thr Phe Ala
Asp Glu Thr 305 310 315
320 Val Asp Lys Asp Ser Val Asn Pro Gly Asp Gln Trp Phe Glu Lys Met
325 330 335 Ile Ser Gly Met
Tyr Leu Gly Glu Ile Val Arg Leu Val Leu Ala Arg 340
345 350 Met Ala Lys Glu Ala Glu Leu Phe Gly
Gly Asn Val Pro Val Lys Leu 355 360
365 Leu Glu Arg Leu Thr Leu Gly Thr Pro His Val Ser Lys Ile
His Leu 370 375 380
Asp Asn Ser Pro Asp Leu Asp Val Val Ala Lys Val Leu Lys Asp Val 385
390 395 400 Phe Glu Ile Glu Thr
Thr Thr Leu Glu Glu Arg Lys Ile Val His Glu 405
410 415 Val Cys Asp Ile Met Gly Glu Arg Gly Gly
Arg Leu Ala Ala Ala Gly 420 425
430 Leu Tyr Gly Ile Leu Lys Lys Ile Gly Arg Thr Gly Lys Ser Arg
Asn 435 440 445 Gly
Ser Lys Lys Lys Thr Val Ile Ala Met Asp Gly Gly Leu Phe Glu 450
455 460 His His Val Arg Tyr Arg
Ser Tyr Met Glu Glu Ala Leu Gln Glu Leu 465 470
475 480 Met Gly Ser Asp Ala Ala Tyr Glu Val Ala Leu
Arg Leu Gln Asn Asp 485 490
495 Gly Ser Gly Ile Gly Ala Ala Leu Leu Ala Ala Ser His Ser His Phe
500 505 510 Lys
752762DNAPhyscomitrella patens 75gttttttgtc atgggctctg ctctgctgcc
tttgtggcga agtggcgcag ctggcttctt 60cttctgcata cttcgcagga agcgctcgaa
gctccacgaa tattgagaga gagctggaga 120agaagaagga attggtacta ctactactac
tactactact actacggtga caatgccaat 180gccagaaacg tccactcttc gggacatagc
gcttgtggta aaatctgagg cgctgtcgag 240gaggagaatt gagtctgtga tttgcaggga
gaagtggtga ggaagtgagc gatctgaggg 300ggagatgatt ttattttttg gtttgttcct
cgtgtgagga gtgagtggtg aagggggtag 360agagagagag cagagaagag gagaggtttt
gagtgagttt gagagagaga gagagagaga 420gagagagaag cggggaaaga tggcgcaatc
gaaagccaga gtgggagtgt gcattgcctg 480tgcagctgcg acgtgcgctg tagcagctgt
gattgtggcg cggcgcgtga agtttcattc 540tcagaagtgt gctgcgcgga aaattctggt
ggagtttcag gaggcatgcg acacgtctct 600gctgcggttg cgcatggtgg tggatgctat
ggcggctgag atgcacgctg gtcttgtttc 660ggaaggcggt agcactctca aaatgcttcc
tacctacatt gatcgcttgc cagacgggaa 720tgagcgagga ctttattatg ctgtggattt
aggtggcacc aacttccgag ttctccgtgt 780tcagctgggt gggttagagg ggagggtaat
caaccaagag tatgaggaag tgcccatccc 840tcctcatgtg atgcttggaa caagtaaaca
attatttgat tttattgcca aggaattggt 900gagctttgtg gcaagagaag gtcaggactt
caggctacat gcaggtcagc agcgggaaat 960agggtttacc ttctcatttc ctgtggacca
aacggcagta aatagtggta aacttttgca 1020gtggaccaag gggttcaaag tgaacgatgc
tattggccag gatgtggttg cagcacttca 1080gaggagcatt gaatccttag ggcacaagat
gaggatatct gctttgatca atgatactgt 1140aggaacgcta gccggcggtc gctactggaa
taatgacgtg atgataggtg ttattttggg 1200taccgggacc aatgcctgtt acgtggagcg
agctgaagct gtatcgaagt ggggtggcga 1260gattccgaag tctggacaga tggtgataaa
catggagtgg gggaatttct ggtcatcaca 1320tttgcctagg acttatgtgg atgaatcctt
ggataacgag agtttgaacc caggagaata 1380cggttttgaa aagatgatct ctggaatgta
tttgggcgat tgtgtgaggc gcgtgcttgt 1440tagaatggca caacaagcag gcatatttgg
tccccgtgtt ccacacaggc ttttggaagc 1500tttctcactt aagacaccgg atatgtcaaa
aatgcatcag gataataata acgatctaag 1560agtggttgga gaaattctaa acagtgttta
ccagattcaa aacaccacat tgggaattcg 1620aaagattgta gtggaggttt gcgatgtggt
gtgtaagaga ggtgctagac tggcaggtgc 1680gggaattgta ggaatattga agaaaattgg
aagggatgga agtgcggcga atggggttat 1740caagcgtaac ctgtttgaac agagtgacat
gaatggttac catgacgacg accctatgca 1800atatacatca gacgtgaaaa ccgttgttgc
tatagacggt ggtttgtatg aacactacac 1860caagttccga gaatacatgc aagatgctgt
gtttgaactt cttggagaag catcaaagaa 1920tgtctccata cagctttcca aagatggatc
aggcattgga gcagcccttc ttgctgcatc 1980gcatgccgag catctttctt cttgataacg
atgaaaccaa ctacagtctt gtgaaatatg 2040agtctttgct gattgaaacc tcttagttct
aatgttagaa atgtgtatac caatccgtca 2100agggggtgcg agttagcttc ctttggagcc
cttggtttat cggctcgcca tttctgtaga 2160aaggttcgct ttttttaatc attaatcacg
catctcggca gctcatgttt ccaagagtaa 2220cttgggacaa cagatcccct ggctgcatac
gtaagcttat ttgaatacta tggagcaact 2280actcaaaaat ctatttgctt ttgtaaacac
cttaaatcaa aggtgacgct ctttgtctgg 2340tccagcctag agcccaatag tctagcctaa
acctctgatg gctaaagttt ggacgttatg 2400gttttgagca ccaaacgttc tgcactgttt
gcaatgctgc ttttctgtct ttcaatttgt 2460tgtggaagga ccaacaatgc ctgtagatac
gctgggtctt cattgtccaa gtgaaacccc 2520aaagagctcg ttgaggtttc tattttagtg
aatgctgttg tcgtcaatgt gggctagttg 2580aagtcccaaa ccgcagtatg aggatgcttg
cttgcttgct atattttgag aatatggtat 2640gtagcatcct ctttgctctg ccaaagggtt
acttgtcttc aggatcttcc ttagcagttt 2700aaagaatttt ggattttgtt aactaaagta
gctttcatac ttaaatgcct acttcaaaag 2760ac
276276521PRTPhyscomitrella patens 76Met
Ala Gln Ser Lys Ala Arg Val Gly Val Cys Ile Ala Cys Ala Ala 1
5 10 15 Ala Thr Cys Ala Val Ala
Ala Val Ile Val Ala Arg Arg Val Lys Phe 20
25 30 His Ser Gln Lys Cys Ala Ala Arg Lys Ile
Leu Val Glu Phe Gln Glu 35 40
45 Ala Cys Asp Thr Ser Leu Leu Arg Leu Arg Met Val Val Asp
Ala Met 50 55 60
Ala Ala Glu Met His Ala Gly Leu Val Ser Glu Gly Gly Ser Thr Leu 65
70 75 80 Lys Met Leu Pro Thr
Tyr Ile Asp Arg Leu Pro Asp Gly Asn Glu Arg 85
90 95 Gly Leu Tyr Tyr Ala Val Asp Leu Gly Gly
Thr Asn Phe Arg Val Leu 100 105
110 Arg Val Gln Leu Gly Gly Leu Glu Gly Arg Val Ile Asn Gln Glu
Tyr 115 120 125 Glu
Glu Val Pro Ile Pro Pro His Val Met Leu Gly Thr Ser Lys Gln 130
135 140 Leu Phe Asp Phe Ile Ala
Lys Glu Leu Val Ser Phe Val Ala Arg Glu 145 150
155 160 Gly Gln Asp Phe Arg Leu His Ala Gly Gln Gln
Arg Glu Ile Gly Phe 165 170
175 Thr Phe Ser Phe Pro Val Asp Gln Thr Ala Val Asn Ser Gly Lys Leu
180 185 190 Leu Gln
Trp Thr Lys Gly Phe Lys Val Asn Asp Ala Ile Gly Gln Asp 195
200 205 Val Val Ala Ala Leu Gln Arg
Ser Ile Glu Ser Leu Gly His Lys Met 210 215
220 Arg Ile Ser Ala Leu Ile Asn Asp Thr Val Gly Thr
Leu Ala Gly Gly 225 230 235
240 Arg Tyr Trp Asn Asn Asp Val Met Ile Gly Val Ile Leu Gly Thr Gly
245 250 255 Thr Asn Ala
Cys Tyr Val Glu Arg Ala Glu Ala Val Ser Lys Trp Gly 260
265 270 Gly Glu Ile Pro Lys Ser Gly Gln
Met Val Ile Asn Met Glu Trp Gly 275 280
285 Asn Phe Trp Ser Ser His Leu Pro Arg Thr Tyr Val Asp
Glu Ser Leu 290 295 300
Asp Asn Glu Ser Leu Asn Pro Gly Glu Tyr Gly Phe Glu Lys Met Ile 305
310 315 320 Ser Gly Met Tyr
Leu Gly Asp Cys Val Arg Arg Val Leu Val Arg Met 325
330 335 Ala Gln Gln Ala Gly Ile Phe Gly Pro
Arg Val Pro His Arg Leu Leu 340 345
350 Glu Ala Phe Ser Leu Lys Thr Pro Asp Met Ser Lys Met His
Gln Asp 355 360 365
Asn Asn Asn Asp Leu Arg Val Val Gly Glu Ile Leu Asn Ser Val Tyr 370
375 380 Gln Ile Gln Asn Thr
Thr Leu Gly Ile Arg Lys Ile Val Val Glu Val 385 390
395 400 Cys Asp Val Val Cys Lys Arg Gly Ala Arg
Leu Ala Gly Ala Gly Ile 405 410
415 Val Gly Ile Leu Lys Lys Ile Gly Arg Asp Gly Ser Ala Ala Asn
Gly 420 425 430 Val
Ile Lys Arg Asn Leu Phe Glu Gln Ser Asp Met Asn Gly Tyr His 435
440 445 Asp Asp Asp Pro Met Gln
Tyr Thr Ser Asp Val Lys Thr Val Val Ala 450 455
460 Ile Asp Gly Gly Leu Tyr Glu His Tyr Thr Lys
Phe Arg Glu Tyr Met 465 470 475
480 Gln Asp Ala Val Phe Glu Leu Leu Gly Glu Ala Ser Lys Asn Val Ser
485 490 495 Ile Gln
Leu Ser Lys Asp Gly Ser Gly Ile Gly Ala Ala Leu Leu Ala 500
505 510 Ala Ser His Ala Glu His Leu
Ser Ser 515 520 772530DNAPhyscomitrella
patens 77gtgagtgagt gagtgtgtgt gtgagtgtga gagtgttgag ttgtgttgtc
gccgctgagt 60ctggttgtgt tgctcacgcg ctgcagagtc cgggttagcg agagagagag
agagagcgag 120agagctggaa gcgagagaga gaaggctcag atccttggca atgtaagcga
ggctgacatg 180gcggatgtgt ttcaagcgag gttccacaaa cataactcaa atccggaggg
ggagaatttg 240gtgcaagaag cactgggaaa gttcagttta agccgcggag attaagttca
gttgttgcgg 300gtgtcgaggg tgtcgagggt gtcgagggta atggtgtttg gttcggtcga
tcgtctgtgg 360attaggaagg aagagatact ggcgatctgt gaaggtgaat ggttcgggtt
gtgaattgtg 420ttaagcaggg gagggtgttg ttgttgttgt tgtggtgtgt ttgagagagc
gagagagagg 480cagaaatggg acaatcgaag gtattggtag gtgtgtacat tgcttgcgca
gctgcggcat 540gcgcaaccgc agctgtggcg gtgacgcagc gactgaaagt gcgagcgcag
aagtgcactg 600cgcggaaaat tttggtggag tttcaggagg cttgcgagac gcctctgccg
cggttgcggc 660aagttgtgga tgctatggcc gtcgagatgc acgctggcct cgtgtcggaa
ggcggaagca 720agcttaagat gctgcccacc ttcattgatc gcttgccgaa tgggagcgag
aagggccttt 780attatgctgt ggacttgggt gggacaaact tccgggtgct ccgtgtccag
ttgggtggat 840tagaaggtag agtaatcaag caagagtacg aggaagttgc cattccccct
gagctaatgc 900ttggaacaag cgaacaacta tttcatttta ttgccaagga gttggctggc
tttgtggcaa 960gagaaggtga ggaattcagg ttaggcgacg gtcagtcacg ggaaataggg
ttcacctttt 1020ccttcccctg taagcaaacc gctgtaaatt ctgggactct cttgcaatgg
accaagggct 1080tcaaagtgaa cgatgcaatc ggccaagatg tggtcgcagc tcttcaaaag
tgcattgaac 1140gactagggtg caagatgagg atcgctgctt tggttaacga tacagtgggt
actctagctg 1200gtggtcgcta ttggaataac gacgtgatga tagctgttat tttgggtact
ggtaccaatg 1260cctgctacgt tgagcgggct gaatctatat cgaagtggac tggcgagctt
ccgaagtctg 1320gccagatggt gatcaatatg gagtggggaa acttttggtc atcacatttg
cctcggacct 1380atgtggatga gttattggat agcgagagcc tccatccagg agaatatggt
tttgaaaaga 1440tgatttctgg aatgtatttg ggtgattgtg tgaggcgagt gcttgtcagg
atggcacaag 1500aagctggcat cttcggtccc catgttcctc acacactctt ggaatccttc
tcgctccaga 1560caccagaaat gtcaagaatg caccatgatg acagtagcga cctaaaagta
gttgcagaag 1620ttctgaaaag actatatgga atccagaaca caacagtagg gattcgcaaa
atcgtagttg 1680ctgtttgcga cactacgtgc cagcgagggg ccaggctggc agctgctggt
attgtgggaa 1740tactaaagaa aattgggaga gatggaagta cagcaaatgg cttgatgagg
cgaaatgata 1800cgaatggtat ccatgacgag ctctctgtga attctacacc gggaagtggt
aagaccgttg 1860tggctatgga cggtggcttg tatgagcact acagcaagtt ccgaaattac
atgcaagaag 1920ctgtgcgtga gcttctaggg gacgcatcca aaaacgtctc tatagagctc
tccaaggatg 1980gctcaggcat tggggcagcc cttctagctg catcatatgc cgagtatgtg
ccctcctaat 2040acctcttgaa ctattgatta aaattccagt aggtcaaatt gttaggactg
tgctttacca 2100atgcttctat gaggtatgag tctgtttctt ttggaacatt gggttatctg
gctcgtcttg 2160tcttgtataa agcctcgtag ttcaatcatc agataattgt tctccttgtc
agctcataga 2220ctggagagtg acttagggta gtggctttct aggtagcagt caaagtatta
tttgaatact 2280ttgaaaagta tcaaacccat tccttgtgta ggacattacc ttgatacttc
agccacagtt 2340taagaagtct ggtctttttt ttagtgtagt agacgaggtt gtagttaatg
gttgtcagca 2400gaaatgcctt gcagtctttt agcagttgct ggctctttct tgcaatttat
tgctaaacag 2460ccttttgggg ttatactgtc ccttcatctc aatcaaaatc attttggatt
cgcttttccg 2520gacttttggc
253078517PRTPhyscomitrella patens 78Met Gly Gln Ser Lys Val
Leu Val Gly Val Tyr Ile Ala Cys Ala Ala 1 5
10 15 Ala Ala Cys Ala Thr Ala Ala Val Ala Val Thr
Gln Arg Leu Lys Val 20 25
30 Arg Ala Gln Lys Cys Thr Ala Arg Lys Ile Leu Val Glu Phe Gln
Glu 35 40 45 Ala
Cys Glu Thr Pro Leu Pro Arg Leu Arg Gln Val Val Asp Ala Met 50
55 60 Ala Val Glu Met His Ala
Gly Leu Val Ser Glu Gly Gly Ser Lys Leu 65 70
75 80 Lys Met Leu Pro Thr Phe Ile Asp Arg Leu Pro
Asn Gly Ser Glu Lys 85 90
95 Gly Leu Tyr Tyr Ala Val Asp Leu Gly Gly Thr Asn Phe Arg Val Leu
100 105 110 Arg Val
Gln Leu Gly Gly Leu Glu Gly Arg Val Ile Lys Gln Glu Tyr 115
120 125 Glu Glu Val Ala Ile Pro Pro
Glu Leu Met Leu Gly Thr Ser Glu Gln 130 135
140 Leu Phe His Phe Ile Ala Lys Glu Leu Ala Gly Phe
Val Ala Arg Glu 145 150 155
160 Gly Glu Glu Phe Arg Leu Gly Asp Gly Gln Ser Arg Glu Ile Gly Phe
165 170 175 Thr Phe Ser
Phe Pro Cys Lys Gln Thr Ala Val Asn Ser Gly Thr Leu 180
185 190 Leu Gln Trp Thr Lys Gly Phe Lys
Val Asn Asp Ala Ile Gly Gln Asp 195 200
205 Val Val Ala Ala Leu Gln Lys Cys Ile Glu Arg Leu Gly
Cys Lys Met 210 215 220
Arg Ile Ala Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala Gly Gly 225
230 235 240 Arg Tyr Trp Asn
Asn Asp Val Met Ile Ala Val Ile Leu Gly Thr Gly 245
250 255 Thr Asn Ala Cys Tyr Val Glu Arg Ala
Glu Ser Ile Ser Lys Trp Thr 260 265
270 Gly Glu Leu Pro Lys Ser Gly Gln Met Val Ile Asn Met Glu
Trp Gly 275 280 285
Asn Phe Trp Ser Ser His Leu Pro Arg Thr Tyr Val Asp Glu Leu Leu 290
295 300 Asp Ser Glu Ser Leu
His Pro Gly Glu Tyr Gly Phe Glu Lys Met Ile 305 310
315 320 Ser Gly Met Tyr Leu Gly Asp Cys Val Arg
Arg Val Leu Val Arg Met 325 330
335 Ala Gln Glu Ala Gly Ile Phe Gly Pro His Val Pro His Thr Leu
Leu 340 345 350 Glu
Ser Phe Ser Leu Gln Thr Pro Glu Met Ser Arg Met His His Asp 355
360 365 Asp Ser Ser Asp Leu Lys
Val Val Ala Glu Val Leu Lys Arg Leu Tyr 370 375
380 Gly Ile Gln Asn Thr Thr Val Gly Ile Arg Lys
Ile Val Val Ala Val 385 390 395
400 Cys Asp Thr Thr Cys Gln Arg Gly Ala Arg Leu Ala Ala Ala Gly Ile
405 410 415 Val Gly
Ile Leu Lys Lys Ile Gly Arg Asp Gly Ser Thr Ala Asn Gly 420
425 430 Leu Met Arg Arg Asn Asp Thr
Asn Gly Ile His Asp Glu Leu Ser Val 435 440
445 Asn Ser Thr Pro Gly Ser Gly Lys Thr Val Val Ala
Met Asp Gly Gly 450 455 460
Leu Tyr Glu His Tyr Ser Lys Phe Arg Asn Tyr Met Gln Glu Ala Val 465
470 475 480 Arg Glu Leu
Leu Gly Asp Ala Ser Lys Asn Val Ser Ile Glu Leu Ser 485
490 495 Lys Asp Gly Ser Gly Ile Gly Ala
Ala Leu Leu Ala Ala Ser Tyr Ala 500 505
510 Glu Tyr Val Pro Ser 515
792087DNAPhyscomitrella patens 79gactggcgat ctcgcggacg acggttggaa
gggaggttag tagctgccca gggttagaat 60tgcttatgtc gaagtgacag attctatgat
cacaacgact agaagagctt tgtaggattt 120taacgaactc tgtttttctc tcttattctt
tctctgtctt caatcttttt ttcaagttgt 180gcagccgatc ggttgagtgg tttggggatt
atcttgcagc ttcagacgat tgcaacatgg 240aaaatttaaa gaattcggaa tcttgcgatc
caagctcttt cctacggcat tttcggaaat 300cttccgctac gcctgtgatg ctattgcgtc
atattgctca ggccatggcg accgagatgc 360aagagggtct tgaccatcct ggcgagcgca
agctgaaaat gctccccacg tatctcgaat 420gcttgcctac agggaatgaa agagggttgt
tttacgccat tgacttggga ggtactaact 480tccgagtatt gcgagtacaa ctagatggga
aggaagggcg catcctgaag caggaatcca 540tacaagttcc cattcctcaa gaagtaatga
ctggaagcag taaggatctt tttggcttcc 600ttgccaagac tattgttcaa ttcgtatcca
gagaagcaga ccttggcttt gagtgtgtag 660cactcaatca gaaacgggat ataggcttca
cgttctcatt tcctgtcaat caaacaaaag 720tcaacggagg ctctatcaac gcatggacca
aaggcttttc catcagtgat ggggttggtg 780aagatgtagt cgatcaacta gagatagctc
ttgcagatat gggctcggtg aatacgaaag 840ttgtgtgctt ggttaatgat acagtaggga
ctttggcaca gtgcagatac tggaacgatg 900atgcaatggt gggcgtcatt ttgggcacag
gatcaaatgc ctgctatgtt gagcgtgctg 960cagccatatc gtgctggagc agtcctccag
aagccgatga tttgacggtt gtaaatattg 1020aatggggcaa tttttgttct gaacttcttc
ctcggacgtt tgccgatgag gggttggatg 1080cagatagttt gaatcctggt caacaggaat
tcgagaaaat gatcggcggt atgtacttgg 1140gtgaaattct tcgccgtgtg ctgctcaaga
tggccgaaga tacaggcctg ttcggctcgg 1200aaattccaga aagactgacg aagcctttca
gccttctgac tcctcacatg tcgacaatgc 1260acggtgatga tacttctagc ttggaagtgg
tgggttctgt gattgaagaa gccattgggg 1320tgaaatatac aacgctagct acacggaaag
tcgtgtacga cgtgtgtgac atcattgcgg 1380aaagaggtgc tcgactctca gctgcaggga
ttgtgggaat cctgacgaag attaacagat 1440gtggagattt aatgctcagt tgcctcacta
caccaaacga cactgaagta aaaaagacag 1500ttattgccat agacggtagt ttatatgaga
agtaccctaa gtttcggaac tacatggaag 1560acgccatgaa ggagatgttg ggcgaagact
atgccaacaa tgtgacaact acactatcca 1620aagacgggtc gggagttgga gccgcgcttc
ttgcagccgc tatttctact gagctacgat 1680tgggcgacgt tggagtcacc aagtgataga
aaggacattg ttagatacat cctgtacatt 1740gcgtaaaggt caggcaccct ctaatctgaa
caatgcttta gttgagtgaa agtgagcgcc 1800attttacacc gtctaggatt gaacatgcca
gagctgtttg agcgggaaaa aattggtcag 1860agttctgatg accacaggaa agaagtccag
catgtaatgt gggttaatgc gcaatggtta 1920catcaaacag gactgtttat gtacgtaatg
tgcacttcca gctgcggatt ttcagctgaa 1980gtgcagtcac agctgatggc ctacatcaca
aaagtgaacc cttgaagtta ccgcaagtat 2040taatgttgaa gcttgagtat ctaactcagc
aagtatcttc ttctctt 208780489PRTPhyscomitrella patens
80Met Glu Asn Leu Lys Asn Ser Glu Ser Cys Asp Pro Ser Ser Phe Leu 1
5 10 15 Arg His Phe Arg
Lys Ser Ser Ala Thr Pro Val Met Leu Leu Arg His 20
25 30 Ile Ala Gln Ala Met Ala Thr Glu Met
Gln Glu Gly Leu Asp His Pro 35 40
45 Gly Glu Arg Lys Leu Lys Met Leu Pro Thr Tyr Leu Glu Cys
Leu Pro 50 55 60
Thr Gly Asn Glu Arg Gly Leu Phe Tyr Ala Ile Asp Leu Gly Gly Thr 65
70 75 80 Asn Phe Arg Val Leu
Arg Val Gln Leu Asp Gly Lys Glu Gly Arg Ile 85
90 95 Leu Lys Gln Glu Ser Ile Gln Val Pro Ile
Pro Gln Glu Val Met Thr 100 105
110 Gly Ser Ser Lys Asp Leu Phe Gly Phe Leu Ala Lys Thr Ile Val
Gln 115 120 125 Phe
Val Ser Arg Glu Ala Asp Leu Gly Phe Glu Cys Val Ala Leu Asn 130
135 140 Gln Lys Arg Asp Ile Gly
Phe Thr Phe Ser Phe Pro Val Asn Gln Thr 145 150
155 160 Lys Val Asn Gly Gly Ser Ile Asn Ala Trp Thr
Lys Gly Phe Ser Ile 165 170
175 Ser Asp Gly Val Gly Glu Asp Val Val Asp Gln Leu Glu Ile Ala Leu
180 185 190 Ala Asp
Met Gly Ser Val Asn Thr Lys Val Val Cys Leu Val Asn Asp 195
200 205 Thr Val Gly Thr Leu Ala Gln
Cys Arg Tyr Trp Asn Asp Asp Ala Met 210 215
220 Val Gly Val Ile Leu Gly Thr Gly Ser Asn Ala Cys
Tyr Val Glu Arg 225 230 235
240 Ala Ala Ala Ile Ser Cys Trp Ser Ser Pro Pro Glu Ala Asp Asp Leu
245 250 255 Thr Val Val
Asn Ile Glu Trp Gly Asn Phe Cys Ser Glu Leu Leu Pro 260
265 270 Arg Thr Phe Ala Asp Glu Gly Leu
Asp Ala Asp Ser Leu Asn Pro Gly 275 280
285 Gln Gln Glu Phe Glu Lys Met Ile Gly Gly Met Tyr Leu
Gly Glu Ile 290 295 300
Leu Arg Arg Val Leu Leu Lys Met Ala Glu Asp Thr Gly Leu Phe Gly 305
310 315 320 Ser Glu Ile Pro
Glu Arg Leu Thr Lys Pro Phe Ser Leu Leu Thr Pro 325
330 335 His Met Ser Thr Met His Gly Asp Asp
Thr Ser Ser Leu Glu Val Val 340 345
350 Gly Ser Val Ile Glu Glu Ala Ile Gly Val Lys Tyr Thr Thr
Leu Ala 355 360 365
Thr Arg Lys Val Val Tyr Asp Val Cys Asp Ile Ile Ala Glu Arg Gly 370
375 380 Ala Arg Leu Ser Ala
Ala Gly Ile Val Gly Ile Leu Thr Lys Ile Asn 385 390
395 400 Arg Cys Gly Asp Leu Met Leu Ser Cys Leu
Thr Thr Pro Asn Asp Thr 405 410
415 Glu Val Lys Lys Thr Val Ile Ala Ile Asp Gly Ser Leu Tyr Glu
Lys 420 425 430 Tyr
Pro Lys Phe Arg Asn Tyr Met Glu Asp Ala Met Lys Glu Met Leu 435
440 445 Gly Glu Asp Tyr Ala Asn
Asn Val Thr Thr Thr Leu Ser Lys Asp Gly 450 455
460 Ser Gly Val Gly Ala Ala Leu Leu Ala Ala Ala
Ile Ser Thr Glu Leu 465 470 475
480 Arg Leu Gly Asp Val Gly Val Thr Lys 485
812177DNAPhyscomitrella patens 81gcgattaagg gaagaagcaa ccggagcgat
gtcgatgacg gtggagcgcg tttcgctctt 60ttcgcgaggg gcgggggtga cgatgcatgg
cctgctgagc gcgagggtgc aatgcgtgag 120gtctccatgg tgggggctgc ccaaagtcgc
tggggcgacg ccgaatgcga acagggtggt 180gcgagtgcac aatgcagatc ggttggtgca
cgggttgcgg ctggccgctg cgactccgct 240cccgctgctg cgccaagtgg cggatgctct
ggtgggagaa atgtgcgctg ggctggagga 300ggaagggggc agcgatcagc tcaagatgct
gccatcgtac gtcgagaatt tgcccactgg 360ggatgaggaa gggctgtttt acgctgtaga
ccttggtggc acaaattttc gggtattgcg 420attacatttg ggcgggaaag gccaagttct
gagccaagag tccaaggaga ttgccatacc 480tcgcgaactt atggtgggca ctggcaagga
tcttttcgac ttcattgcca atacgcttgc 540cacatttgtc gacacggagg acattctgct
tgattcaaag tccaacaagc acagggaagc 600cgggtttgcg ttttcttttc ccgttcgtca
aacatcagta aaatctggca atgtcattca 660atggaccaaa ggctttaaaa tcgatgatgc
gataggcaaa gacattgtga agcagttcca 720ggatgcgatt agtcgcagtg gtcacgatgt
tgaaatttct gccttggtga acgatactgt 780cggtacatta gccggaggta ggtataactt
tcaggaagag acaatgatcg gttgcatact 840tggcacaggg acaaatgctt gttacgtgga
acgagctgat gctgtgaaga aatggaaaga 900ggcacttccc aaatcagggg agatggtcat
caacttagag tggggaaatt ttcgatcacc 960ttggctgccg cgcacatttg ctgatgatga
ggtcgacaaa gaaagtgtaa accccggaga 1020ccagtggttt gaaaaaatgg tgtcaggcat
gtaccttggc gaaatcgtac gacacatgct 1080gctgaggctt gctgaagagg caacgttgtt
tggggacact gtccccgaaa aactcagaga 1140acaacagtct cttgaaacca aacatgtttc
taaaatccat gctgacattt catcggagct 1200ccaaacagta gcgactgttc ttcacgaagt
tctcaggatc catgacacca ctctcgaaca 1260gcggaggatt gtgcacagct tatgcgacat
ggttggccag cggggtggaa ggttggctgc 1320tgcaggactc tacgggatac tgaaaaaaat
tggtagagct ggaccaaaca agaatggctt 1380cgccttatct cgacagaaga agacgacggt
ggtggcaatg gatggaggtc tttatgaaca 1440ccaccacccg taccggaaat acatggagga
cgctctccaa gagttggtcg gcacaaacgg 1500accctacgag gtgtttttaa gactacaaaa
tgacggttca ggaattggag cggcattgct 1560tgccgcatct cactctcgac acagacaaac
cgagtagcga ggcagacgac tcgtcatctg 1620gctacctctg tagagtggtg tgaataccgc
gaatgtgttc ccgaaaagcc agcaatttct 1680acttagactt catcatatct gggaataagg
agctgaactg accaggaaat caacaatttt 1740ccctttggaa ttcctttttg atcggctctt
gtggcggttg catcgcggtg aagacgatag 1800aggtaatcct ctacctcggc aagcattgat
atcttccagc acctacacta gtcatatgtc 1860gcacctgctg gataagatca gagtaacgtg
tagaggggtt tggactttac actggaggcg 1920tagcatggct gtagacgggg cgttgagtgc
ttctacctgt ccagctaggt aagtaaattg 1980tatctctctg agacgtgtct tggttgtgct
acacagatca gaagaggtct gcatagacaa 2040ttgatggaca catgtaaacc cgtagaacaa
ggcatcgtca actctttcta gcaacctaat 2100ccaaataatt actatctaga tttgttcgcg
aacatttgta aaatatactt ttaaacgatg 2160gaggaaaagt caaaaac
217782522PRTPhyscomitrella patens 82Met
Ser Met Thr Val Glu Arg Val Ser Leu Phe Ser Arg Gly Ala Gly 1
5 10 15 Val Thr Met His Gly Leu
Leu Ser Ala Arg Val Gln Cys Val Arg Ser 20
25 30 Pro Trp Trp Gly Leu Pro Lys Val Ala Gly
Ala Thr Pro Asn Ala Asn 35 40
45 Arg Val Val Arg Val His Asn Ala Asp Arg Leu Val His Gly
Leu Arg 50 55 60
Leu Ala Ala Ala Thr Pro Leu Pro Leu Leu Arg Gln Val Ala Asp Ala 65
70 75 80 Leu Val Gly Glu Met
Cys Ala Gly Leu Glu Glu Glu Gly Gly Ser Asp 85
90 95 Gln Leu Lys Met Leu Pro Ser Tyr Val Glu
Asn Leu Pro Thr Gly Asp 100 105
110 Glu Glu Gly Leu Phe Tyr Ala Val Asp Leu Gly Gly Thr Asn Phe
Arg 115 120 125 Val
Leu Arg Leu His Leu Gly Gly Lys Gly Gln Val Leu Ser Gln Glu 130
135 140 Ser Lys Glu Ile Ala Ile
Pro Arg Glu Leu Met Val Gly Thr Gly Lys 145 150
155 160 Asp Leu Phe Asp Phe Ile Ala Asn Thr Leu Ala
Thr Phe Val Asp Thr 165 170
175 Glu Asp Ile Leu Leu Asp Ser Lys Ser Asn Lys His Arg Glu Ala Gly
180 185 190 Phe Ala
Phe Ser Phe Pro Val Arg Gln Thr Ser Val Lys Ser Gly Asn 195
200 205 Val Ile Gln Trp Thr Lys Gly
Phe Lys Ile Asp Asp Ala Ile Gly Lys 210 215
220 Asp Ile Val Lys Gln Phe Gln Asp Ala Ile Ser Arg
Ser Gly His Asp 225 230 235
240 Val Glu Ile Ser Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala Gly
245 250 255 Gly Arg Tyr
Asn Phe Gln Glu Glu Thr Met Ile Gly Cys Ile Leu Gly 260
265 270 Thr Gly Thr Asn Ala Cys Tyr Val
Glu Arg Ala Asp Ala Val Lys Lys 275 280
285 Trp Lys Glu Ala Leu Pro Lys Ser Gly Glu Met Val Ile
Asn Leu Glu 290 295 300
Trp Gly Asn Phe Arg Ser Pro Trp Leu Pro Arg Thr Phe Ala Asp Asp 305
310 315 320 Glu Val Asp Lys
Glu Ser Val Asn Pro Gly Asp Gln Trp Phe Glu Lys 325
330 335 Met Val Ser Gly Met Tyr Leu Gly Glu
Ile Val Arg His Met Leu Leu 340 345
350 Arg Leu Ala Glu Glu Ala Thr Leu Phe Gly Asp Thr Val Pro
Glu Lys 355 360 365
Leu Arg Glu Gln Gln Ser Leu Glu Thr Lys His Val Ser Lys Ile His 370
375 380 Ala Asp Ile Ser Ser
Glu Leu Gln Thr Val Ala Thr Val Leu His Glu 385 390
395 400 Val Leu Arg Ile His Asp Thr Thr Leu Glu
Gln Arg Arg Ile Val His 405 410
415 Ser Leu Cys Asp Met Val Gly Gln Arg Gly Gly Arg Leu Ala Ala
Ala 420 425 430 Gly
Leu Tyr Gly Ile Leu Lys Lys Ile Gly Arg Ala Gly Pro Asn Lys 435
440 445 Asn Gly Phe Ala Leu Ser
Arg Gln Lys Lys Thr Thr Val Val Ala Met 450 455
460 Asp Gly Gly Leu Tyr Glu His His His Pro Tyr
Arg Lys Tyr Met Glu 465 470 475
480 Asp Ala Leu Gln Glu Leu Val Gly Thr Asn Gly Pro Tyr Glu Val Phe
485 490 495 Leu Arg
Leu Gln Asn Asp Gly Ser Gly Ile Gly Ala Ala Leu Leu Ala 500
505 510 Ala Ser His Ser Arg His Arg
Gln Thr Glu 515 520
831569DNAPhyscomitrella patens 83atgagagcaa tggcgatcgg gtcggtgctg
ggttgtgtcg ggttgcagtt ctccgcagtg 60ccgacgtttc atggatcggg ggtggcgagt
ccgaagatga ggacgcaatg cagagcttgg 120cggagaacta tgtccatgtc ggttcagcag
ccgtcgaaac gggtccagcg tgcagagacg 180ttgctgcatg acttccgtcg gtcgtccgcc
actccccttc ctcttctgca cctggtggcg 240gatgctctcg tgcacgagat gtacgctgga
ttggtctcag aaggggggag tgatcagctg 300aagatgctcc caacgtacgt cgaggagttg
ccttctggga gtgaaaaggg gctgttctac 360gccgtggact tgggcgggac aaactttcgc
gttctaaggg tgcaattagg cggccacact 420ggtgagattt tgagtcagga gttcaaggag
gtggccatac ctccagagct catggtgggc 480accggcaagg atcttttcga cttcattgcg
ggtacgctcg cgtcatttgt cgacaccgaa 540gacgagtcat tgaaagccca ctttgctcag
tcgggcaaag tcagggaatc cggcttcgcg 600ttctcctttc ctgttcgcca gacgtctgtc
aaatccggca ttgtcatcca ctggaccaag 660ggcttcaaag ttgacgatgc ggtgggcaaa
gatatcgtga aacagttcca ggatgcaatc 720agcaggagtg gtcatcagat tgcgatttca
gccctggtga acgacaccgt cggtaccttg 780gccggaggca ggttcaactt tgaggaggag
accatgattg gctgcatcat tgggacaggc 840acgaacgcgt gctacgtgga acgtgctgat
agtgttcaaa agtgggcgga tccactgccg 900aaatcaggac agatggtgat caacatggaa
tggggtaatt ttcattcacc cttcctgcca 960cgcacatttg ctgacgacat tgttgacaaa
gacagcgtaa accctggaga ccagtggttt 1020gagaaaatga tatctgggat gtaccttggc
gagatcgtgc gtctcgtgct tgcgaggatg 1080gctgaagaag cgcagttgtt tggtggcagc
ccccccgcca agctgttgga gaaactcagc 1140ctcggcaccc cacatgtttc gaagatgcat
gctgacgctt caccggattt gcaagtcgtt 1200gccgaagttc tggaggacgt ctacgggatc
gaaaccacca cgctcgagga gaggaagatt 1260gtacgcgagg tgtgtgacat cttgggcaaa
cgaggaggaa ggctagctgc agcaggtctt 1320tacggcatac tgaagaagat aggcagaacg
gagagatctc agaacggatt ccaacaaaag 1380aagaagacgg tgattgcaat ggacggaggt
ctgttcgagc atcacgagcc ttaccgggcc 1440tacatggagg aggcactcca cgagttgatg
ggctccgaag ccctttacga ggtgtcttta 1500aggctgcaaa acgatgggtc cggtgtaggt
gctgcgttgc ttgctgcttc ccattcacaa 1560tttaaatag
156984522PRTPhyscomitrella patens 84Met
Arg Ala Met Ala Ile Gly Ser Val Leu Gly Cys Val Gly Leu Gln 1
5 10 15 Phe Ser Ala Val Pro Thr
Phe His Gly Ser Gly Val Ala Ser Pro Lys 20
25 30 Met Arg Thr Gln Cys Arg Ala Trp Arg Arg
Thr Met Ser Met Ser Val 35 40
45 Gln Gln Pro Ser Lys Arg Val Gln Arg Ala Glu Thr Leu Leu
His Asp 50 55 60
Phe Arg Arg Ser Ser Ala Thr Pro Leu Pro Leu Leu His Leu Val Ala 65
70 75 80 Asp Ala Leu Val His
Glu Met Tyr Ala Gly Leu Val Ser Glu Gly Gly 85
90 95 Ser Asp Gln Leu Lys Met Leu Pro Thr Tyr
Val Glu Glu Leu Pro Ser 100 105
110 Gly Ser Glu Lys Gly Leu Phe Tyr Ala Val Asp Leu Gly Gly Thr
Asn 115 120 125 Phe
Arg Val Leu Arg Val Gln Leu Gly Gly His Thr Gly Glu Ile Leu 130
135 140 Ser Gln Glu Phe Lys Glu
Val Ala Ile Pro Pro Glu Leu Met Val Gly 145 150
155 160 Thr Gly Lys Asp Leu Phe Asp Phe Ile Ala Gly
Thr Leu Ala Ser Phe 165 170
175 Val Asp Thr Glu Asp Glu Ser Leu Lys Ala His Phe Ala Gln Ser Gly
180 185 190 Lys Val
Arg Glu Ser Gly Phe Ala Phe Ser Phe Pro Val Arg Gln Thr 195
200 205 Ser Val Lys Ser Gly Ile Val
Ile His Trp Thr Lys Gly Phe Lys Val 210 215
220 Asp Asp Ala Val Gly Lys Asp Ile Val Lys Gln Phe
Gln Asp Ala Ile 225 230 235
240 Ser Arg Ser Gly His Gln Ile Ala Ile Ser Ala Leu Val Asn Asp Thr
245 250 255 Val Gly Thr
Leu Ala Gly Gly Arg Phe Asn Phe Glu Glu Glu Thr Met 260
265 270 Ile Gly Cys Ile Ile Gly Thr Gly
Thr Asn Ala Cys Tyr Val Glu Arg 275 280
285 Ala Asp Ser Val Gln Lys Trp Ala Asp Pro Leu Pro Lys
Ser Gly Gln 290 295 300
Met Val Ile Asn Met Glu Trp Gly Asn Phe His Ser Pro Phe Leu Pro 305
310 315 320 Arg Thr Phe Ala
Asp Asp Ile Val Asp Lys Asp Ser Val Asn Pro Gly 325
330 335 Asp Gln Trp Phe Glu Lys Met Ile Ser
Gly Met Tyr Leu Gly Glu Ile 340 345
350 Val Arg Leu Val Leu Ala Arg Met Ala Glu Glu Ala Gln Leu
Phe Gly 355 360 365
Gly Ser Pro Pro Ala Lys Leu Leu Glu Lys Leu Ser Leu Gly Thr Pro 370
375 380 His Val Ser Lys Met
His Ala Asp Ala Ser Pro Asp Leu Gln Val Val 385 390
395 400 Ala Glu Val Leu Glu Asp Val Tyr Gly Ile
Glu Thr Thr Thr Leu Glu 405 410
415 Glu Arg Lys Ile Val Arg Glu Val Cys Asp Ile Leu Gly Lys Arg
Gly 420 425 430 Gly
Arg Leu Ala Ala Ala Gly Leu Tyr Gly Ile Leu Lys Lys Ile Gly 435
440 445 Arg Thr Glu Arg Ser Gln
Asn Gly Phe Gln Gln Lys Lys Lys Thr Val 450 455
460 Ile Ala Met Asp Gly Gly Leu Phe Glu His His
Glu Pro Tyr Arg Ala 465 470 475
480 Tyr Met Glu Glu Ala Leu His Glu Leu Met Gly Ser Glu Ala Leu Tyr
485 490 495 Glu Val
Ser Leu Arg Leu Gln Asn Asp Gly Ser Gly Val Gly Ala Ala 500
505 510 Leu Leu Ala Ala Ser His Ser
Gln Phe Lys 515 520
852446DNAPhyscomitrella patens 85gttttatagc aaagagtgtg tgtgtgggcg
tgcatgtatg tgtgtgtgtg cttctgtgct 60tggaagtgaa gcgtgctgga agcattccaa
ccttcacgag attgagagcc ggaaggaagg 120gattggaacc gattgatggc aatgtgagtg
gcgcctggcc atggcggatg cgttcaaggc 180gaggtacagc agcggagctc agattcggac
agggaaaatt cggagcggtg agggggagga 240aaattatttg tagagggaga ggagtgattt
cggaggaagg aggtcattga ttgattgatt 300gattgattga ttgcgagact ttttagaaga
ggtgaggagg cgaagaaagt gagcttttat 360ttttatttgt gtgtgtgtga gagagaggga
ggggtagaga cagaggagag gaaaaatgac 420acaatcgaag gtaatgacgg gcgtgtacat
cgcctgcgca gctgcggcgt gcgctgctgc 480ggctgtgatt gtatcacggc gcttgaaggt
tcgatcacag aaatgcactg cgcggaaaat 540tctgctggag tttcaggagg cctgttacac
gcctttggcg cgcctgcgcc aggtggtgga 600tgctatggcg gtcgagatgc acgctggtct
tgtttcggaa ggtggaagca aacttaagat 660gcttcccacc tacattgatc gcttgcctga
cgggcatgag aggggtctat actatgctgt 720ggatttgggt ggtaccaact tccgagtact
ccgcgttcaa ttgggtggac tggagggtag 780ggtgatcaaa caagagtatg aggaagtggc
tattcctcct gagctcatgc ttggaacaag 840tgaacagttg tttgatttca ttgccaagga
gttggtgagt tttgtagcaa gggaaggtca 900ggacttcaga ttgcatgctg gtcaaaatcg
agaaatagga tttacttttt cattccccgt 960gaagcaaact gcagtgaact ctggcactct
tctgcagtgg actaaagggt tcaaagtgaa 1020cgatgctgtt ggcgaggatg tagttgcagc
gcttcagagg ggtattgagc gaagagggta 1080caagatgagg attgctgctt tggtcaacga
taccgtagga accctagctg gcggacggta 1140ttggaacaat gacgtaatga taggtgttat
cttgggtact gggaccaacg cctgctatgt 1200ggagagagct gaagctgtat cgaagtgggc
tggcgacatt ccaaagtccg gggagatggt 1260tataaacatg gagtggggga atttctggtc
atctcacttg cctcggacct atgtggatga 1320gtcattggac aacgagagtt tgaatccagg
agaatatggt ttcgagaaaa tgatctctgg 1380aatgtatttg ggtgattgtg tgaggcgcgt
gcttgttaga atggcacaag aagcctgcat 1440ttttggcacc cctgtcccac acaagctttt
ggaagcattt tctcttatga ctccagacat 1500gtcgaaaatg catcatgatg atagttctga
tttaaaggtg gttgctgaag ttctgaaaag 1560agtttacggg atccagaaca ctacagtagg
aatccgcaaa atcgttgttg cagtttgtga 1620tacggtgtgc cagcgagggg caagactagc
tgctgctggc attgtgggaa tattgaagaa 1680aattggaaga gatggaagtg cggcaaatgg
tgttattaag cggaatacct tcgaacagag 1740tgatatgaat ggttttcatg acgaagttcc
tgtgcattac acatcgggcg gcagaactgt 1800tgtggctatg gacggtggtt tgtatgaaca
ctacaccaag tttcgaaact acatgcaaga 1860agctgtggtc gagctcctag gagaaggatc
taagaacgtc gtcattgagc tttctaaaga 1920cggatcaggc attggagcag ctcttcttgc
tgcatcacat gcagagtacg tcatttcctg 1980ataatgatga gaaaaactac attgtcttat
gaaacctgag gctttgttga ttagaaattc 2040ttagttctga tcgtgagggc tgtgtacttg
actaatgcat caaggatgta cgagtttgct 2100tcttttgaag cacatggttt gttggctcgt
catttctcct gtataaagtc tcttcagtct 2160tcatcattag ttatcctcat agtttcgaga
gtcactcagg acgttacatt tcttggtaac 2220aaattaggtt ttatttgagt actttggagc
aagtactcaa gcgattttct tctgtacatc 2280gtcttgaaac aaaggtcact tcttaagtct
ggtccagcat agagcggaat agttctgtac 2340taatcaacct ctggtaccca agggttggag
gcaatgcttt tcagcataaa atgtttgcag 2400tcttctactg cgactgttca attctaaact
tctgttttgg aaggtc 244686521PRTPhyscomitrella patens
86Met Thr Gln Ser Lys Val Met Thr Gly Val Tyr Ile Ala Cys Ala Ala 1
5 10 15 Ala Ala Cys Ala
Ala Ala Ala Val Ile Val Ser Arg Arg Leu Lys Val 20
25 30 Arg Ser Gln Lys Cys Thr Ala Arg Lys
Ile Leu Leu Glu Phe Gln Glu 35 40
45 Ala Cys Tyr Thr Pro Leu Ala Arg Leu Arg Gln Val Val Asp
Ala Met 50 55 60
Ala Val Glu Met His Ala Gly Leu Val Ser Glu Gly Gly Ser Lys Leu 65
70 75 80 Lys Met Leu Pro Thr
Tyr Ile Asp Arg Leu Pro Asp Gly His Glu Arg 85
90 95 Gly Leu Tyr Tyr Ala Val Asp Leu Gly Gly
Thr Asn Phe Arg Val Leu 100 105
110 Arg Val Gln Leu Gly Gly Leu Glu Gly Arg Val Ile Lys Gln Glu
Tyr 115 120 125 Glu
Glu Val Ala Ile Pro Pro Glu Leu Met Leu Gly Thr Ser Glu Gln 130
135 140 Leu Phe Asp Phe Ile Ala
Lys Glu Leu Val Ser Phe Val Ala Arg Glu 145 150
155 160 Gly Gln Asp Phe Arg Leu His Ala Gly Gln Asn
Arg Glu Ile Gly Phe 165 170
175 Thr Phe Ser Phe Pro Val Lys Gln Thr Ala Val Asn Ser Gly Thr Leu
180 185 190 Leu Gln
Trp Thr Lys Gly Phe Lys Val Asn Asp Ala Val Gly Glu Asp 195
200 205 Val Val Ala Ala Leu Gln Arg
Gly Ile Glu Arg Arg Gly Tyr Lys Met 210 215
220 Arg Ile Ala Ala Leu Val Asn Asp Thr Val Gly Thr
Leu Ala Gly Gly 225 230 235
240 Arg Tyr Trp Asn Asn Asp Val Met Ile Gly Val Ile Leu Gly Thr Gly
245 250 255 Thr Asn Ala
Cys Tyr Val Glu Arg Ala Glu Ala Val Ser Lys Trp Ala 260
265 270 Gly Asp Ile Pro Lys Ser Gly Glu
Met Val Ile Asn Met Glu Trp Gly 275 280
285 Asn Phe Trp Ser Ser His Leu Pro Arg Thr Tyr Val Asp
Glu Ser Leu 290 295 300
Asp Asn Glu Ser Leu Asn Pro Gly Glu Tyr Gly Phe Glu Lys Met Ile 305
310 315 320 Ser Gly Met Tyr
Leu Gly Asp Cys Val Arg Arg Val Leu Val Arg Met 325
330 335 Ala Gln Glu Ala Cys Ile Phe Gly Thr
Pro Val Pro His Lys Leu Leu 340 345
350 Glu Ala Phe Ser Leu Met Thr Pro Asp Met Ser Lys Met His
His Asp 355 360 365
Asp Ser Ser Asp Leu Lys Val Val Ala Glu Val Leu Lys Arg Val Tyr 370
375 380 Gly Ile Gln Asn Thr
Thr Val Gly Ile Arg Lys Ile Val Val Ala Val 385 390
395 400 Cys Asp Thr Val Cys Gln Arg Gly Ala Arg
Leu Ala Ala Ala Gly Ile 405 410
415 Val Gly Ile Leu Lys Lys Ile Gly Arg Asp Gly Ser Ala Ala Asn
Gly 420 425 430 Val
Ile Lys Arg Asn Thr Phe Glu Gln Ser Asp Met Asn Gly Phe His 435
440 445 Asp Glu Val Pro Val His
Tyr Thr Ser Gly Gly Arg Thr Val Val Ala 450 455
460 Met Asp Gly Gly Leu Tyr Glu His Tyr Thr Lys
Phe Arg Asn Tyr Met 465 470 475
480 Gln Glu Ala Val Val Glu Leu Leu Gly Glu Gly Ser Lys Asn Val Val
485 490 495 Ile Glu
Leu Ser Lys Asp Gly Ser Gly Ile Gly Ala Ala Leu Leu Ala 500
505 510 Ala Ser His Ala Glu Tyr Val
Ile Ser 515 520 871829DNAPhyscomitrella
patens 87actggttcgt gttccagtgt tgacgttcga tgaagtcgat gttttgagga
ttcgaagggt 60aatacattgg cgatctgcgg gaggtgagtg ttttgtgagg gttggagttg
aggcaaggag 120agaaggaagg aaggcaggaa gggagaacag ttttgtgagg aggagagaga
tggggcaatc 180gaaagcaatg gcgggggtgt atatcgcctg tgcagctgcg gcgtgcgctg
ctgcagctgt 240ggtgttgacg cagcgagtga aagtccgatc gcaaaagtac acagcgcgga
aaattttagt 300ggagtttcag gaggcttgtg agacgcctct gccgcggttg aggcaggttg
tggatgctat 360ggcagtcgag atgcatgctg gtcttgtttc ggaaggtggt agcaagctta
agatgctgcc 420tacatttatt gaccacttgc cagacgggaa tgagaaaggc ctttattatg
ctgtggattt 480gggtggcaca aacttccggg tgctccgtac ccaattgggt gggctggagg
gtagagtaat 540taaacaagaa tatgaggagg ttgccattcc tcctgagctg atgcttggaa
caagcgaaca 600attgtttgat tttattgcca aggagttggt tagctttgtg gcaagagaag
gtgaggattt 660tagattgcac gaaggtcagt cacgggagat agggttcacc ttttcctttc
cctgtaagca 720aaccgctgtg aactctggta ctctcttgca gtggaccaag ggctttaaag
tcaacgatgc 780aatcggccag gatgtggttg cagctctaca agggagcatt gaacggcgag
ggtacaagat 840gaggattgct gctttggtta acgacacggt aggaactcta gctggtggtc
gctattggaa 900taacgacgtc atgatagctg ttattttggg tactggcacc aacgcctgct
atgtggagcg 960agctgaatct atatcgaagt ggggtggcga gcttccgaag tctggccaga
tggtgatcaa 1020tatggagtgg ggaaattttt ggtcgtcaca tttgcctcgg acctacgtgg
acgagctatt 1080ggataacgag agtctgaatc caggagaata cggcttcgag aaaatgattt
cgggaatgta 1140tttgggtgat tgtgtgaggc gtgtacttgt tagaatggct cagcaagctg
gcatcttcgg 1200tccccatgtt ccacacaagc tcttggaagc cttcacactt caaacaccag
atatgtcgaa 1260aatgcatcat gatgacagta gtgacctcaa aatggttgca gaagttctga
agacagtata 1320tgggatccat aacacaacac tggggattcg caaaattgta cttgctgttt
gcgatattgt 1380gtgccagcga ggggccagat tagcagctgc tggtattgtg ggaatattaa
agaaaattgg 1440aagggatgga agtacagcaa atggttttat caggcgaaat gacgtgaatg
gtattcacga 1500tgagctcacc gtgaattcca ttgggggaag cgggaaaacc gtcgtggcta
tggacggtgg 1560tttgtatgag cactacagta agttccggaa ctacatgcaa caagctgtac
gtgaactcct 1620aggggacgca gccaaaaacg tcttcataga gctttcgaag gatggctcag
gcattggagc 1680agccattctt gctgcatcac atgccgagta tgtaccaacc taatatcaac
ccgataccgc 1740ttaaattaat aacatttaat tatggaaccc aaggccttgt tgattaaaac
cctttagttc 1800ttatcgttag gactgtactc tacttatgc
182988517PRTPhyscomitrella patens 88Met Gly Gln Ser Lys Ala
Met Ala Gly Val Tyr Ile Ala Cys Ala Ala 1 5
10 15 Ala Ala Cys Ala Ala Ala Ala Val Val Leu Thr
Gln Arg Val Lys Val 20 25
30 Arg Ser Gln Lys Tyr Thr Ala Arg Lys Ile Leu Val Glu Phe Gln
Glu 35 40 45 Ala
Cys Glu Thr Pro Leu Pro Arg Leu Arg Gln Val Val Asp Ala Met 50
55 60 Ala Val Glu Met His Ala
Gly Leu Val Ser Glu Gly Gly Ser Lys Leu 65 70
75 80 Lys Met Leu Pro Thr Phe Ile Asp His Leu Pro
Asp Gly Asn Glu Lys 85 90
95 Gly Leu Tyr Tyr Ala Val Asp Leu Gly Gly Thr Asn Phe Arg Val Leu
100 105 110 Arg Thr
Gln Leu Gly Gly Leu Glu Gly Arg Val Ile Lys Gln Glu Tyr 115
120 125 Glu Glu Val Ala Ile Pro Pro
Glu Leu Met Leu Gly Thr Ser Glu Gln 130 135
140 Leu Phe Asp Phe Ile Ala Lys Glu Leu Val Ser Phe
Val Ala Arg Glu 145 150 155
160 Gly Glu Asp Phe Arg Leu His Glu Gly Gln Ser Arg Glu Ile Gly Phe
165 170 175 Thr Phe Ser
Phe Pro Cys Lys Gln Thr Ala Val Asn Ser Gly Thr Leu 180
185 190 Leu Gln Trp Thr Lys Gly Phe Lys
Val Asn Asp Ala Ile Gly Gln Asp 195 200
205 Val Val Ala Ala Leu Gln Gly Ser Ile Glu Arg Arg Gly
Tyr Lys Met 210 215 220
Arg Ile Ala Ala Leu Val Asn Asp Thr Val Gly Thr Leu Ala Gly Gly 225
230 235 240 Arg Tyr Trp Asn
Asn Asp Val Met Ile Ala Val Ile Leu Gly Thr Gly 245
250 255 Thr Asn Ala Cys Tyr Val Glu Arg Ala
Glu Ser Ile Ser Lys Trp Gly 260 265
270 Gly Glu Leu Pro Lys Ser Gly Gln Met Val Ile Asn Met Glu
Trp Gly 275 280 285
Asn Phe Trp Ser Ser His Leu Pro Arg Thr Tyr Val Asp Glu Leu Leu 290
295 300 Asp Asn Glu Ser Leu
Asn Pro Gly Glu Tyr Gly Phe Glu Lys Met Ile 305 310
315 320 Ser Gly Met Tyr Leu Gly Asp Cys Val Arg
Arg Val Leu Val Arg Met 325 330
335 Ala Gln Gln Ala Gly Ile Phe Gly Pro His Val Pro His Lys Leu
Leu 340 345 350 Glu
Ala Phe Thr Leu Gln Thr Pro Asp Met Ser Lys Met His His Asp 355
360 365 Asp Ser Ser Asp Leu Lys
Met Val Ala Glu Val Leu Lys Thr Val Tyr 370 375
380 Gly Ile His Asn Thr Thr Leu Gly Ile Arg Lys
Ile Val Leu Ala Val 385 390 395
400 Cys Asp Ile Val Cys Gln Arg Gly Ala Arg Leu Ala Ala Ala Gly Ile
405 410 415 Val Gly
Ile Leu Lys Lys Ile Gly Arg Asp Gly Ser Thr Ala Asn Gly 420
425 430 Phe Ile Arg Arg Asn Asp Val
Asn Gly Ile His Asp Glu Leu Thr Val 435 440
445 Asn Ser Ile Gly Gly Ser Gly Lys Thr Val Val Ala
Met Asp Gly Gly 450 455 460
Leu Tyr Glu His Tyr Ser Lys Phe Arg Asn Tyr Met Gln Gln Ala Val 465
470 475 480 Arg Glu Leu
Leu Gly Asp Ala Ala Lys Asn Val Phe Ile Glu Leu Ser 485
490 495 Lys Asp Gly Ser Gly Ile Gly Ala
Ala Ile Leu Ala Ala Ser His Ala 500 505
510 Glu Tyr Val Pro Thr 515
891929DNAPhyscomitrella patens 89gctcttgata atggctggtg gactatgacg
atattctgat gttacgattg cggtcctaga 60gtagtaaatc aagaagggag gattcaatta
tggaaaggaa catggaggtc ctctagcgag 120caacaagctt gtgcaggata gcttgtcgag
aacatcccct gatggcagat aataaggcgt 180ggattaacgc cctcacagca ggtacggcag
tggttgctgt tgccactgtt gtttgtgtgt 240ggcagaggat gcaggggagc acacaaacaa
cgagaactac tcgccaccaa gcgctcgggc 300ttttacatga atttcagcat gctgcagcga
cccccttgta cgttttgcga cagatctcgg 360agcacatggc ggtagagatg cgtgccggcc
taatgacaga gggcaagagc agcctgctca 420tgctgccaac tttcgtggaa aatcttccgg
acggaaatga gacaggtctg ttctacgcct 480tggatttagg cggaaccaac tttcgggttc
tgcgctgtct ccttggggga agagaaggca 540gagtactgaa acaggagtac gaggaggtcc
ctattcccaa aatactcatg tttggcacaa 600gtgaggaact tttcgacttc atagccaaga
agctggtcga cttcgtcaac cgagaggggg 660atgagtacaa gccgcgcggt ggccgacaac
aaacagtccg cgaactcggg ctgacgtttt 720ctttccctgt gaagcaaaca tcagtaagat
ctggtgttct catccagtgg agcaaaggct 780tcttagttgc tgacggggtt ggagcagatg
tggtggcgct gttgcagcgt gcgatcaatc 840gccagcatgg accaaagatt gaggtggttg
tgctggtgaa cgacacggtg ggcactttgg 900caggagggcg ctattggaac gaagatgcga
tggttgggat gattctgggg acaggtacca 960acgcttgcta tgtggagcgt gatctgcctg
cccatgccaa ttccagcacg ggtgaaatgg 1020ttatcaacat ggagtgggct gggttctggt
cgtcacatct gccacgcact tatgccgatg 1080agcagcttga cagtgagagc gtcaaccctg
gtcaagcggg attcgagaaa atgattggag 1140gcatgtactt gggagaaatc gtccgtcgtg
tattgttcaa aatggcgggg gaggctgcgt 1200tgttcgggga tgaaataccg cacaaattga
aagagccgtt tgtcttaatg acttcggaga 1260tgtcgaaaat gcatgctgat gaatccagcg
acttacgggt ggtgggcaca attttgagag 1320atgtgtttgg gattcaaaaa accgagttgc
ctacgcgtag gattgtgcac gacgtgtgcg 1380acactgtaac cttgcgcagt gcaaggctgg
cggcagctgg aattgtgggc atcttcaaga 1440aaattggcgg ggactcgtgg gactccccac
atctgagcgg gacctccaaa gcagtccacc 1500accactcccg agtgactagc acaggagtag
gagccggaaa gaccatggtg gccatggacg 1560gaggactctt cgagcattac atccagtacc
ggatctacat gcaagccgct gttagtgagc 1620tactgagtga agccgcagcg agtcgcctgg
tgatccagtt ggctaaagat ggatctggta 1680ttggagcgag cattcttgca gcctgtcact
ccaagtacag gtaacaggta ctcgatcagt 1740catatgctga gctgatgatg cttttccgga
gctgagtgct cagccaattg aatctctatg 1800ctttgcaaca cacccccaca tcgcagacag
tcaaatttct ttttggtcgt ggtgctgaat 1860atcataagta gttggcaaaa cgaatgtgcg
tttctcattc accaagagag tgcgttatcc 1920atccttgaa
192990520PRTPhyscomitrella patens 90Met
Ala Asp Asn Lys Ala Trp Ile Asn Ala Leu Thr Ala Gly Thr Ala 1
5 10 15 Val Val Ala Val Ala Thr
Val Val Cys Val Trp Gln Arg Met Gln Gly 20
25 30 Ser Thr Gln Thr Thr Arg Thr Thr Arg His
Gln Ala Leu Gly Leu Leu 35 40
45 His Glu Phe Gln His Ala Ala Ala Thr Pro Leu Tyr Val Leu
Arg Gln 50 55 60
Ile Ser Glu His Met Ala Val Glu Met Arg Ala Gly Leu Met Thr Glu 65
70 75 80 Gly Lys Ser Ser Leu
Leu Met Leu Pro Thr Phe Val Glu Asn Leu Pro 85
90 95 Asp Gly Asn Glu Thr Gly Leu Phe Tyr Ala
Leu Asp Leu Gly Gly Thr 100 105
110 Asn Phe Arg Val Leu Arg Cys Leu Leu Gly Gly Arg Glu Gly Arg
Val 115 120 125 Leu
Lys Gln Glu Tyr Glu Glu Val Pro Ile Pro Lys Ile Leu Met Phe 130
135 140 Gly Thr Ser Glu Glu Leu
Phe Asp Phe Ile Ala Lys Lys Leu Val Asp 145 150
155 160 Phe Val Asn Arg Glu Gly Asp Glu Tyr Lys Pro
Arg Gly Gly Arg Gln 165 170
175 Gln Thr Val Arg Glu Leu Gly Leu Thr Phe Ser Phe Pro Val Lys Gln
180 185 190 Thr Ser
Val Arg Ser Gly Val Leu Ile Gln Trp Ser Lys Gly Phe Leu 195
200 205 Val Ala Asp Gly Val Gly Ala
Asp Val Val Ala Leu Leu Gln Arg Ala 210 215
220 Ile Asn Arg Gln His Gly Pro Lys Ile Glu Val Val
Val Leu Val Asn 225 230 235
240 Asp Thr Val Gly Thr Leu Ala Gly Gly Arg Tyr Trp Asn Glu Asp Ala
245 250 255 Met Val Gly
Met Ile Leu Gly Thr Gly Thr Asn Ala Cys Tyr Val Glu 260
265 270 Arg Asp Leu Pro Ala His Ala Asn
Ser Ser Thr Gly Glu Met Val Ile 275 280
285 Asn Met Glu Trp Ala Gly Phe Trp Ser Ser His Leu Pro
Arg Thr Tyr 290 295 300
Ala Asp Glu Gln Leu Asp Ser Glu Ser Val Asn Pro Gly Gln Ala Gly 305
310 315 320 Phe Glu Lys Met
Ile Gly Gly Met Tyr Leu Gly Glu Ile Val Arg Arg 325
330 335 Val Leu Phe Lys Met Ala Gly Glu Ala
Ala Leu Phe Gly Asp Glu Ile 340 345
350 Pro His Lys Leu Lys Glu Pro Phe Val Leu Met Thr Ser Glu
Met Ser 355 360 365
Lys Met His Ala Asp Glu Ser Ser Asp Leu Arg Val Val Gly Thr Ile 370
375 380 Leu Arg Asp Val Phe
Gly Ile Gln Lys Thr Glu Leu Pro Thr Arg Arg 385 390
395 400 Ile Val His Asp Val Cys Asp Thr Val Thr
Leu Arg Ser Ala Arg Leu 405 410
415 Ala Ala Ala Gly Ile Val Gly Ile Phe Lys Lys Ile Gly Gly Asp
Ser 420 425 430 Trp
Asp Ser Pro His Leu Ser Gly Thr Ser Lys Ala Val His His His 435
440 445 Ser Arg Val Thr Ser Thr
Gly Val Gly Ala Gly Lys Thr Met Val Ala 450 455
460 Met Asp Gly Gly Leu Phe Glu His Tyr Ile Gln
Tyr Arg Ile Tyr Met 465 470 475
480 Gln Ala Ala Val Ser Glu Leu Leu Ser Glu Ala Ala Ala Ser Arg Leu
485 490 495 Val Ile
Gln Leu Ala Lys Asp Gly Ser Gly Ile Gly Ala Ser Ile Leu 500
505 510 Ala Ala Cys His Ser Lys Tyr
Arg 515 520 911557DNAPhyscomitrella patens
91atgccggaat tggagaatag gatatggatg aacatgctca tagcaggtgc tgcggtggct
60gccattacta ccgtggtgat tgcctggaat agtacacaga aggcccctct cgtgaaaact
120cgcaaccggg cactagagct tttatacgag tttcagcatg cggctgcaac tcctttacac
180gctttgcggc agattgcgga gcacatggcc attgagatgc gtgcaggact gaggaggggc
240ggcaagagca accttctcat gcttcctact tacgtggaga atcttccgga cggagacgag
300gaaggactgt tctatgcttt ggatttgggg ggtaccaact ttcgggtcct gcgctgtctg
360cttggaggga aagatggcag ggtattgaaa caggaatttg aggaggtctc catacccaaa
420gcgctgatgt tgggcacaag tgcggagctc ttcgacttca tagcggagag actggtcgat
480tttgtgagcc gagaaggaga agggtttaag acgagaaacg gcatgcaaca aacagttcgc
540gaaatgggtt taacgttctc atttcctgtc aaacaacaat ctgtgaaatc gggggccata
600attcagtgga gcaagggctt cgacattgct gatggggttg gagcagatgt cgtggcgctg
660ttgcaaagtg caataaaccg ccagcgtgga ccgaagattg aggtggctgt cttggtcaat
720gacacggtgg gtactctggc tggtgggcgc tactggaacg aagatgtgat ggtgggaatg
780attttgggga caggcaccaa cgcttgctac gtagagcatg atttgcccag ccacgtccaa
840tctggatcgg gcgagatgat aatcaacatg gagtggggtg ggttctggtc gtcacatttg
900ccacggactt tcgccgatga gcagcttgac aaggagagcc tcaaccccgg tcaggcgggt
960tacgaaaaaa tgatcggcgg tatgtacctg ggagaaatag ttcgtcgagt gttgttgagg
1020atggaaaagg aggcttcact atttggtggt ccagtacctt ctaaattaaa agagcccttc
1080agtttgataa ctccagagat tgccaagatg catgccgatg agtcaaagaa cttgcgggtg
1140gtggccaaag ttttgaggga cgtgtttggg gttcaaaaaa ccgatttggc agcaaggagg
1200atcgtgcacg atgtttgtga cattgtaatc atgcgcagtg caaggttggc ggcagcggga
1260atcgtgggca ttttcaaaaa aattgggggc gaggctttcg actcgacgga ttcaaggaca
1320ctcacaaatc tgcactggga ctcccaagag cctcaacaac ttgacacaaa acgaattgtg
1380gtggccgtgg acggagggct gtatgagcac tgcacccaat accgggtcta tatgcgggct
1440gctgtgaatg agcttctgag cgaagcaggg gcaaagcgat tgcaaatcgt gttgtccaag
1500gatgggtcgg gcattggagc aagcatcctt gcagcacgtc actcccagca taggtaa
155792518PRTPhyscomitrella patens 92Met Pro Glu Leu Glu Asn Arg Ile Trp
Met Asn Met Leu Ile Ala Gly 1 5 10
15 Ala Ala Val Ala Ala Ile Thr Thr Val Val Ile Ala Trp Asn
Ser Thr 20 25 30
Gln Lys Ala Pro Leu Val Lys Thr Arg Asn Arg Ala Leu Glu Leu Leu
35 40 45 Tyr Glu Phe Gln
His Ala Ala Ala Thr Pro Leu His Ala Leu Arg Gln 50
55 60 Ile Ala Glu His Met Ala Ile Glu
Met Arg Ala Gly Leu Arg Arg Gly 65 70
75 80 Gly Lys Ser Asn Leu Leu Met Leu Pro Thr Tyr Val
Glu Asn Leu Pro 85 90
95 Asp Gly Asp Glu Glu Gly Leu Phe Tyr Ala Leu Asp Leu Gly Gly Thr
100 105 110 Asn Phe Arg
Val Leu Arg Cys Leu Leu Gly Gly Lys Asp Gly Arg Val 115
120 125 Leu Lys Gln Glu Phe Glu Glu Val
Ser Ile Pro Lys Ala Leu Met Leu 130 135
140 Gly Thr Ser Ala Glu Leu Phe Asp Phe Ile Ala Glu Arg
Leu Val Asp 145 150 155
160 Phe Val Ser Arg Glu Gly Glu Gly Phe Lys Thr Arg Asn Gly Met Gln
165 170 175 Gln Thr Val Arg
Glu Met Gly Leu Thr Phe Ser Phe Pro Val Lys Gln 180
185 190 Gln Ser Val Lys Ser Gly Ala Ile Ile
Gln Trp Ser Lys Gly Phe Asp 195 200
205 Ile Ala Asp Gly Val Gly Ala Asp Val Val Ala Leu Leu Gln
Ser Ala 210 215 220
Ile Asn Arg Gln Arg Gly Pro Lys Ile Glu Val Ala Val Leu Val Asn 225
230 235 240 Asp Thr Val Gly Thr
Leu Ala Gly Gly Arg Tyr Trp Asn Glu Asp Val 245
250 255 Met Val Gly Met Ile Leu Gly Thr Gly Thr
Asn Ala Cys Tyr Val Glu 260 265
270 His Asp Leu Pro Ser His Val Gln Ser Gly Ser Gly Glu Met Ile
Ile 275 280 285 Asn
Met Glu Trp Gly Gly Phe Trp Ser Ser His Leu Pro Arg Thr Phe 290
295 300 Ala Asp Glu Gln Leu Asp
Lys Glu Ser Leu Asn Pro Gly Gln Ala Gly 305 310
315 320 Tyr Glu Lys Met Ile Gly Gly Met Tyr Leu Gly
Glu Ile Val Arg Arg 325 330
335 Val Leu Leu Arg Met Glu Lys Glu Ala Ser Leu Phe Gly Gly Pro Val
340 345 350 Pro Ser
Lys Leu Lys Glu Pro Phe Ser Leu Ile Thr Pro Glu Ile Ala 355
360 365 Lys Met His Ala Asp Glu Ser
Lys Asn Leu Arg Val Val Ala Lys Val 370 375
380 Leu Arg Asp Val Phe Gly Val Gln Lys Thr Asp Leu
Ala Ala Arg Arg 385 390 395
400 Ile Val His Asp Val Cys Asp Ile Val Ile Met Arg Ser Ala Arg Leu
405 410 415 Ala Ala Ala
Gly Ile Val Gly Ile Phe Lys Lys Ile Gly Gly Glu Ala 420
425 430 Phe Asp Ser Thr Asp Ser Arg Thr
Leu Thr Asn Leu His Trp Asp Ser 435 440
445 Gln Glu Pro Gln Gln Leu Asp Thr Lys Arg Ile Val Val
Ala Val Asp 450 455 460
Gly Gly Leu Tyr Glu His Cys Thr Gln Tyr Arg Val Tyr Met Arg Ala 465
470 475 480 Ala Val Asn Glu
Leu Leu Ser Glu Ala Gly Ala Lys Arg Leu Gln Ile 485
490 495 Val Leu Ser Lys Asp Gly Ser Gly Ile
Gly Ala Ser Ile Leu Ala Ala 500 505
510 Arg His Ser Gln His Arg 515
931906DNAPhyscomitrella patens 93aagttgtaaa gtaatctctc cgtgctcggt
tggtttctag aggttaagta tgatatgaga 60gtctcgttga gcattcgatt gtagcatgca
caatcgaact ctaaacaggg atctgccaat 120tagtccaaat cgacgaagaa ataaggttcc
tccctttcga tgattctgat gccttgtagg 180agcagatgac acagatgctg ttaagcacgg
ctgtggcatg tgcaacggtg gcagccgtgg 240cggcagccgt catggtctgg cagaaattcc
acaagcatag tcattgcgat caggcgctcg 300tcttactgta cgaatttcga catgcctgcg
ctaccccttt gtatgtcttg cgccacatct 360cggagcatgt ggctcttgaa atgcaggccg
gtctcaatca gcctggaggc agtcagctca 420tgatgctccc cactttcatt gaaaaactgc
caaatgggtg tgagaagggg cttttctatg 480cattggattt gagtgggacc aattttcgcg
tgttgcgatg ccttttaggt gggcctaatg 540cgcgagtgat aaaacaagag tatgaagtgg
tcgccattcc gcgtgcactc ttgctgggaa 600caagcgagga gctcttcgac ttcatcgcac
agagattgat ctcgtttatt aagctggagg 660gaccggagtt tcagcgggga cataactgga
atggacatca aattcgtgaa ttagggttga 720caatctcttt tcccatctgc cagacctcac
acaacacggg cattctcatc aagtggactg 780aaggattcaa gattgccgac ggggttggca
aagatgtagt ggcaatgctc caaagtgcaa 840tggaccggca gaaaggattt cagattaggg
tggctgtgtt gattaatgac acagttggaa 900caatggcggg tgggcactat tggaatgatg
atgtgatggt aggcgtgatt ctagggacta 960acaccaatgc ctgctacgtt gagtgcaact
tgcccgagga catccagacg aagagcggca 1020agatggtgat ctatatggaa tggggtaggt
tttggtcatc acacctccca cgcacttaca 1080ttgacgagca gcttgacaat gagagtgtga
atccaggcga tagaggattt gagaagatga 1140caggggccat gtacttaggt gagatagttc
gtcgcgttct ggcgaggatg gcacaggaag 1200caaatttatt cggagactcc gtaccaacaa
aattaaaaca acctttcatt ttgttaacac 1260tggagatgtc caaaatgcac gcagatgagt
caccagattt acgaattgtg gacaaagttt 1320tgaaagatgt ctttgatatc aaacggaccg
agttgtcaga gcggaggatt gtgcacagcg 1380tctgcgacac agtgaccatg cgggctgctc
ggctagcagc agcattcatt gtgggtatct 1440tgaagaaaat cgggcgggat ggttgggatg
cgacaggtgt tagtagcaga ctaatggcat 1500taccacgaga ctcggagcat agagccaggc
tcgaattgaa gaggaccgtc gttgctatgg 1560atggaattct atacgagcat tatcaccgct
ttagaatcta catgcaagca gccgtctatg 1620agttgttaag tgaagccgct gccaggaaac
ttgtgattga gctttcgaaa gatgggtcag 1680gtactggagc cagtattctt gctgcttgtc
attcagagtt tgctccctct tattcatgag 1740ttatgactgc cgcatcgaaa ttaattgtgt
cttctttttg tgtaaatatc catcaacgat 1800tggtacaacc cctttggcaa tcaagtgttg
attacccagg tagtattttg ttcacaacct 1860ctttttgaac tttagaatgt gagctatcat
cggtccagta caacat 190694517PRTPhyscomitrella patens
94Met Thr Gln Met Leu Leu Ser Thr Ala Val Ala Cys Ala Thr Val Ala 1
5 10 15 Ala Val Ala Ala
Ala Val Met Val Trp Gln Lys Phe His Lys His Ser 20
25 30 His Cys Asp Gln Ala Leu Val Leu Leu
Tyr Glu Phe Arg His Ala Cys 35 40
45 Ala Thr Pro Leu Tyr Val Leu Arg His Ile Ser Glu His Val
Ala Leu 50 55 60
Glu Met Gln Ala Gly Leu Asn Gln Pro Gly Gly Ser Gln Leu Met Met 65
70 75 80 Leu Pro Thr Phe Ile
Glu Lys Leu Pro Asn Gly Cys Glu Lys Gly Leu 85
90 95 Phe Tyr Ala Leu Asp Leu Ser Gly Thr Asn
Phe Arg Val Leu Arg Cys 100 105
110 Leu Leu Gly Gly Pro Asn Ala Arg Val Ile Lys Gln Glu Tyr Glu
Val 115 120 125 Val
Ala Ile Pro Arg Ala Leu Leu Leu Gly Thr Ser Glu Glu Leu Phe 130
135 140 Asp Phe Ile Ala Gln Arg
Leu Ile Ser Phe Ile Lys Leu Glu Gly Pro 145 150
155 160 Glu Phe Gln Arg Gly His Asn Trp Asn Gly His
Gln Ile Arg Glu Leu 165 170
175 Gly Leu Thr Ile Ser Phe Pro Ile Cys Gln Thr Ser His Asn Thr Gly
180 185 190 Ile Leu
Ile Lys Trp Thr Glu Gly Phe Lys Ile Ala Asp Gly Val Gly 195
200 205 Lys Asp Val Val Ala Met Leu
Gln Ser Ala Met Asp Arg Gln Lys Gly 210 215
220 Phe Gln Ile Arg Val Ala Val Leu Ile Asn Asp Thr
Val Gly Thr Met 225 230 235
240 Ala Gly Gly His Tyr Trp Asn Asp Asp Val Met Val Gly Val Ile Leu
245 250 255 Gly Thr Asn
Thr Asn Ala Cys Tyr Val Glu Cys Asn Leu Pro Glu Asp 260
265 270 Ile Gln Thr Lys Ser Gly Lys Met
Val Ile Tyr Met Glu Trp Gly Arg 275 280
285 Phe Trp Ser Ser His Leu Pro Arg Thr Tyr Ile Asp Glu
Gln Leu Asp 290 295 300
Asn Glu Ser Val Asn Pro Gly Asp Arg Gly Phe Glu Lys Met Thr Gly 305
310 315 320 Ala Met Tyr Leu
Gly Glu Ile Val Arg Arg Val Leu Ala Arg Met Ala 325
330 335 Gln Glu Ala Asn Leu Phe Gly Asp Ser
Val Pro Thr Lys Leu Lys Gln 340 345
350 Pro Phe Ile Leu Leu Thr Leu Glu Met Ser Lys Met His Ala
Asp Glu 355 360 365
Ser Pro Asp Leu Arg Ile Val Asp Lys Val Leu Lys Asp Val Phe Asp 370
375 380 Ile Lys Arg Thr Glu
Leu Ser Glu Arg Arg Ile Val His Ser Val Cys 385 390
395 400 Asp Thr Val Thr Met Arg Ala Ala Arg Leu
Ala Ala Ala Phe Ile Val 405 410
415 Gly Ile Leu Lys Lys Ile Gly Arg Asp Gly Trp Asp Ala Thr Gly
Val 420 425 430 Ser
Ser Arg Leu Met Ala Leu Pro Arg Asp Ser Glu His Arg Ala Arg 435
440 445 Leu Glu Leu Lys Arg Thr
Val Val Ala Met Asp Gly Ile Leu Tyr Glu 450 455
460 His Tyr His Arg Phe Arg Ile Tyr Met Gln Ala
Ala Val Tyr Glu Leu 465 470 475
480 Leu Ser Glu Ala Ala Ala Arg Lys Leu Val Ile Glu Leu Ser Lys Asp
485 490 495 Gly Ser
Gly Thr Gly Ala Ser Ile Leu Ala Ala Cys His Ser Glu Phe 500
505 510 Ala Pro Ser Tyr Ser
515 952642DNAArabidopsis thaliana 95atattattca cgcttctcgt
aattaatcat ttggtgaatt atataattcg aagtatcttt 60tgacagctct agttcgagat
attatacaat aaaatcaact agctactcta tttattttat 120tgtgttcgat aatgagacat
ttcatgtcgg ttttgtttaa ttttataaga agagacatta 180ttgagcgagt tttctttttg
ggttcctgca tgaacaatgg caaattgtgg gaactttaca 240tatcgcttct ctcagactct
gagcacttct ataaagctac actttctgac caagacacta 300gtctgactct ttggttcggc
catttccaac cagcaccagt tgctggacca cttgcttctc 360catcttaatg aagccacaaa
tgaacacatt ctcctcaaag cttcctctca aatttttgca 420aaatatttga aaaccatttt
cttgttttcc tattaaaatt tgtaatatat agtaaacttt 480taacagtaat aaagaaaaaa
gattagtaag ttctcttttt taattcatat acgtgtagtc 540ttttttccac gatattttta
ggattgggtc ccaataaaca aatggataaa cggtaagagt 600ctaacgagag gccgtctcta
gataacgtaa gaaagctaca aaaattcaac atgtgaatat 660ttgagggaaa gtttgagtgg
atggacaaag aatataccct ctctattatg aatcatttta 720aggcaaaaaa agggagaaag
atgacaaaga gaagtgagtc aatttaattt aaaacgtata 780aagcaaacaa aagagacact
tgattattct cattcatctg caaaagtaac catatttttt 840aagcaaaatt tctaaatttc
gttatgtagc catttaattc catttaatca tttttggaaa 900cattaaccaa tgatgacaat
ttcttgctgg ttatgaacct ttcccattat ttcctttatt 960tgatgcgttc atcttcacaa
atatttagtc ctcatttagc atttgctttt aattcattgc 1020attaatcttt catgacgaca
acacacaact cttctttgtc ctcattctta tctacatata 1080tcatctaagg tgttatttta
attgttccat tctcactaca aataattttt gcaaaaacgt 1140ttctaatgct ctagtaaata
tttggttttg atccgtgtac gatacacgta tgatgtaatc 1200ctatatcttt atacgacggc
tcgctgattt ttctgggttg aaagatttta aaattgatct 1260tctaataaat cacaacaaat
ttatacatta gacataaatt gagaattact ctttctcttg 1320acaaaaaaaa cattcaaaaa
gcttttggga atcacttttt caataaatca cacatcccaa 1380tagcttcaaa cagttgtagt
aaaatgtacg tcaggttcac ttggtgacat atataattaa 1440gagagaaaaa aaacacatgc
acatcatcat tagcaaagag atatccttgt cacagaccaa 1500aatattttaa agagcacaca
ttgctccaaa ctctcgtcga ctttgataca ttattaagaa 1560ctcttaccaa tttatatgtg
gaatccatct ttccatcaat tataatctta ataaactcta 1620cactcttctc gcagctgatt
tactaataaa aaataaaaat aaaaaaaact ttttgtttcg 1680atgctttctc tcactcggac
aaatactaac gatcatttta gaataagtca accatattga 1740aaacgactct caactaatta
cgtaaactta tattttgaag tagtcattct tacattgttg 1800taaacacttt tccataaatg
catgaagttt ataattataa aatatttaga catattccta 1860tcaatttttt atagatcacg
atttacatga atctccgaaa agagaaacat caaaagaata 1920tggaaaaatt tccgaaataa
aaactctaga agaattgctg aggtcacgcc ggttttggct 1980gtgtagagtg attcgtaaca
tcttttaggc tgaacatgaa tgagcatgtc tccactactt 2040agactcacat aattttagct
ctataaaaga aataattttc ggtgattgtt tgaaattaat 2100atatatcgat catacttttg
tcgtagcgtt tgaacgtatg atcataatgg gtccatttag 2160agatgacgaa tgtatgatga
ttagagtgat gtaaaaggtt taaaaaacta ctacatgaac 2220tccatttctt aaccttaact
ttcttaccat tggtgcgagc cgatccgacg tggaccaatc 2280aaattgtttc tacaaagctg
aaaaaaattt acacactttt gcaacttttc tctcaatcct 2340tttttaaaaa gaagatttag
aaaaggaaaa tagtagtaaa agagagcata aaattcaaca 2400aacccttctg aagaaagtga
cttttcttag ttctattaat actctctctc tctgcagatc 2460tgttacttca ttatctcttc
ctttttttca ttagattcca ttaaccttca aaagtttttc 2520taatacattc tctctgctca
caactttttt ttctttcaat acttgtaaag aaaaaataga 2580gctttcttct tcttctcttt
tactgttagc tttgcacagc attgcagctg tgaataacta 2640aa
2642961694DNAArabidopsis
thaliana 96gctcctgctc ttcctcaaga ccttccttga ttcgccgccg gtatgttctc
cgtctgtggt 60agcgcctttg gaacactcta ccaacgccgc catgaaagga tctctcatgg
ccgcagggga 120cgtgttcttc ttacatctgg tgttagggct atggttactc cagtgaggag
ggagaggcaa 180gaggttgctt aatgattcgt ttttccggtg atacgagaac tctttaggtt
taccgggaag 240cttttcccat gaaaatggga tgccaagtgg atggagagga gttgccggag
agttgccgga 300gaataggagg gaattggagg aggaggaaga gagtgatcgc cgggttgaaa
tgttaaccgt 360cgaggagaat ttgaccgagt tggatcgtct agtaggtaca attcgggtcc
ttggcgaagt 420atccattcaa aatagtgttt agttttggac ttgagaactt gttgtctctt
tgatctcttt 480tatataaaac tttggacgtg taggacaaac ttgtcaacat aagaaacaaa
atggttgcaa 540cagagaggat gaatttataa gttttcaaca ccgcttttct tattagacgg
acaacaatct 600atagtggagt aaatttttat ttttggtaaa atggttagtg aattcaaata
tctaaatttt 660gtgactcact aacattaaca aatatgcata agacataaaa aaaagaaaga
ataattctta 720tgaaacaaga aaaaaaacct atacaatcaa tctttaggaa ttgacgatgt
agaattgtag 780atgataaatt ttctcaaata tagatgggcc taatgaaggg tgccgcttat
tggatctgac 840ccattttgag gacattaata ttttcattgg ttataagcct tttaatcaaa
attgtcatta 900aattgatgtc tccctctcgg gtcattttcc tttctccctc acaattaatg
tagactttag 960caatttgcac gctgtgcttt gtctttatat ttagtaacac aaacattttg
acttgtcttg 1020tagagttttt ctcttttatt tttctatcca atatgaaaac taaaagtgtt
ctcgtataca 1080tatattaaaa ttaaagaaac ctatgaaaac accaatacaa atgcgatatt
gttttcagtt 1140cgacgtttca tgtttgttag aaaatttcta atgacgtttg tataaaatag
acaattaaac 1200gccaaacact acatctgtgt tttcgaacaa tattgcgtct gcgtttcctt
catctatctc 1260tctcagtgtc acaatgtctg aactaagaga cagctgtaaa ctatcattaa
gacataaact 1320accaaagtat caagctaatg taaaaattac tctcatttcc acgtaacaaa
ttgagttagc 1380ttaagatatt agtgaaacta ggtttgaatt ttcttcttct tcttccatgc
atcctccgaa 1440aaaagggaac caatcaaaac tgtttgcata tcaaactcca acactttaca
gcaaatgcaa 1500tctataatct gtgatttatc caataaaaac ctgtgattta tgtttggctc
cagcgatgaa 1560agtctatgca tgtgatctct atccaacatg agtaattgtt cagaaaataa
aaagtagctg 1620aaatgtatct atataaagaa tcatccacaa gtactatttt cacacactac
ttcaaaatca 1680ctactcaaga aata
1694971292DNAArabidopsis thaliana 97catatggtat gatgatatgc
tttgtttctc tgcttctctt actaatttga agctgttgga 60ttgatttgtc tcttcttacg
ttcccttctt ttttttttcg ttttcttttg tcgtatagac 120caggcagggg ctagggccta
gtgatgggta ttggcccaat actattgggt tatttgcctg 180gtttattatt tcgattttag
gttaattcaa ttttaagaat acgtagattt gtttggttta 240gtttggtttg gttgcactaa
gttcggtttt acataaatag aatctaacac tactaattgt 300tatacgtaaa atacaacaac
aataacagat ttttcgtttc aattttcgtt taagagggta 360gacattttgg tttggtttgg
ttcatttttt ttttcccttt caaattcaca tccttcacgt 420agatgacaaa ataaagaaaa
acatgaatga aagttgtaac ttgtaagcat caacatggaa 480atcatatcac aaagaacaca
aatctaacta atgggtcttt tcacatattg gtataattat 540aagttgtaag aatattagtt
aaacagaggc aacgagagat gcgtgatata tgaaaagttg 600aaaacaaaag acatggatct
aaagagtcaa gcaaaatgta atatcttttt ttcttctaaa 660cttgaggatg tccaagttgc
agtgaatgat tccctttaat catggagaaa ttcaatgaaa 720taattgtgtt tcttcccaca
ctttatcttt atttattttc ttaccacaat tacaactatt 780atcacaaaaa tgtaagtaac
atagcttgtg actcttcttc catttatgag ttgattatca 840ctatatttat aagtaattac
caacgaatgt tccaaattaa gcaaaatatt gtaatcgata 900cactatgtat tcatctacaa
tatgttaacg agctcctttt atggaaatat ttcgattgaa 960aaaacatttg atggatcgtt
cactaaataa ataatccagt aacgttttct taagggagat 1020atacatattc gtgtggagat
caacatatct tcgttaattg actacgcaaa atagttaatg 1080gaaaaggcag agtgactcgt
gagcttggca gatccaaaag aggttgtcaa gaaaaagcag 1140atttaaaagt tcttccctct
tctttaagtc acccattaat ttcacatata tgtacataca 1200tgttgcattt aactcatata
catacatatt ctcacatcta taaagagagc ataagactca 1260gagagatcta gaggaagaga
gagagagaaa ga 1292982174DNAVitis vinifera
98gtgattaaat ttttattcat attcttgaat atcccaaatt aatcaagata actcatattt
60gtttaaaaaa aaattaaatt ttaggtttaa aaactaaaat tacctattta cccctctaac
120taaaattgtt tctttgactc ttttaagaga gtagtcaatt tgaaggcacc tccccattca
180ttggtcaaaa ctaatgactt aaatgctttt cacattgctt aaatcaacaa tttttaattt
240aagataatcg tcatactcaa gagcacgaaa caaaaaaaag agtataatgg tgaaactctc
300tctctcctca caattatagt aacacactct aaaaaaacaa aactctaaat cataaaaata
360ttaaatatat aataagagta aaatcgtcgt tgctcaaata aataaacaaa caaaaaattc
420aaaaaacgct actccttcaa cctttgtgag ctgtttggac aacctattcc tcacaagttc
480aacgaaaaac tccaccccta atatttctta caaagcacaa ctaaattaat ttaaacttca
540taaatattca atgttaattt ataacatctg caagaagaat atgcttttat atcatctctc
600tctctctaaa agataacatt cataggaaag taaaaatagt cacataactt ataagaacca
660cattccatta acttggatct tactttaaaa ctattatttc atttttccta attttggtaa
720aattcctcat aaacaagact aatccatccc atcattaaaa ttttcagtgc tttctttgct
780ttctaacgcc aacacccttt ctttattttt aatttttttt ttcttttata gcaaatttaa
840ggaaaagtga aaatttcatt gttcttacac ggaattgctt ttcttcaaaa ctaagacaac
900aaaaattgaa gttaaaaaaa aaaaataaaa atcatgagtg ataatggaat ccaaagatat
960aaataataag aaatatgaga aaatgtttat tttaaaaaag gatttttttt tttaaatgat
1020acctctataa atgtgtgaag aagtttggta agacacctct ataaatacaa catagctcta
1080aaattttaat attaagatgt tttaaaatta aaattctcaa tatctattgc taggattaat
1140tttaagtttc ttacaatgaa atcatagaat aatttaagga aaaaaagcta aagttttgag
1200taagagacaa attaattttt tcatagcgtt aattaagtac ttaagtctta ttttcatata
1260tttcttatga acacaaaaaa aagttttaaa aaaaaaaatt tatctagtaa ttaataatat
1320ttttatacat gtatttctat tttaatttct aacaaatttg aacttattca gacttcaaat
1380aaaaaaacaa tctccacctt ctaactcacg tggtccaata tttatcataa gatagtagtc
1440tcatgcttag atgtttcatc tgaatacaaa aaagtcaaaa attttaactt gatttttatt
1500taaatttcaa acaaacccta acccattcat acttcgaata ttaaacaata aataaaagaa
1560aaattttcat atttgatgaa accaaatttg atgcatgaga tttcaatgtg tgacctgatc
1620ccaagatagt aaaaaattga attcagatct caatcttctt ccaaattaac taaattaaaa
1680tatatatata aaagagaata taaaatgttg tgtccatgaa atgatgaata gcatggctta
1740aatgcatggt gtcaacccac ctaaacccat tactgaaata agaggcttag aagaagtgaa
1800ataaatacga tgactatgaa gtttcctatt agctttgttg gtgaagtgct gtggcatgtg
1860agtgtacaac agaaaagaga ctcacagaag gaattcaact ccatacttct gactttggtt
1920ctctctgtgg atatcaacca agctatattt ataataaagt tgagaaaaag gcaaggtaga
1980tcaatggaaa tgggtgaaaa aaagagaggt tttatgagtc actcttagta tataaacacc
2040ctcacatggt gatatgctac tatttgaaag gcacgaagag atagatagat agagagagag
2100agagagagag acagagagag agagagagag agagagagag agagagagag agagtttggg
2160atgaaaacaa gcca
2174993001DNAArabidopsis thaliana 99aaccaaaaaa tcgggttagg gcgggttaaa
attttaggaa atttccttat tgggtagagt 60tttactaaac ccgtggatat ccgattggac
cggaaattat ccgttatcta aaaagagtat 120tcaaaaaccc aaacattaat ttaatatcca
aaatattaat tatatgatat tattttattt 180gattttaaat atatagtaaa ctgcgagttg
tatatgtttt cttgatatta tttatattgt 240ttagtgttta aaattataca cttgtatttt
gattgttaat tttagagttt cacctgtagt 300ataccatctt atattaatat cgatttaaac
ccgtcaattc taggattttc cagcttgtat 360taaaaattga atcacatcat acacataaaa
aaatctaata tgttattaat tattgttgta 420tataagatta taaattctta aaataatatg
catgaaattg aatataaata tttaaattat 480gacccagtac ttagtaataa attttcttaa
atctattttt gacccgttat aatatttttt 540catgtattga acagtttata ttcgttttta
aaagtttaaa ttatggcata tgcgaaaaaa 600ctctaattat ttttttataa cgatgatatt
attttttcgc aaaaatagaa tcatataaag 660atgagaggtg aactataata attaataaaa
aattaatatg ataatttaga tatcaaatct 720tatttgttga ttttaattgg ttaatttttt
gaaaattaat aatgtatttc gttttttaat 780gaaatttaat taattaaatt agtatttgac
tttttaattt ttaaagaaat gaattaattt 840actctttaaa ttttatttct aatggcatac
ctatgtaatt acttacaaaa aataaggtta 900catttaaaat gtacttccca aataatatag
aaggatgtga aactagataa aaaaacgttg 960tattatcggt cagatttttc ttttacgttt
tatttatttt cctttttatt aacgttttcg 1020tcatgtttca acttcaggca gttacaactt
acaacagttt ttaacaaaat agaaataaat 1080tagaaagcga tttccagcta cacaagtaac
aaaaatttga agattatact taaaaagaga 1140gagagatatt catctcccca aatttaaaga
gacatttcaa aattgaacta gcgaatgcaa 1200taaatactag gaaatatatt cgtagctcct
ttaccctatg ataattaata ttcagtcatc 1260tactattcta ctttctttgt tattataaga
agaagtagaa ttacattaaa atctgtgaac 1320atttgagtgt taaataaagt gaagcttcgt
taagtttacc taccattcct atttctttgt 1380ttttcatgac attttactag ttgaatgcta
accaaaattc tttgatcaga tctaaactaa 1440atataatatt tgtaaccgta aaatataatg
tgaagagagt actgtacata gtgaatatat 1500tcatattcct tttcaagaaa atactagaac
attcatacac atctttaaaa attactagat 1560tctagtcact tggcataagc aaaccaagct
atccagccaa ataattagtg ttgggctagt 1620ggtgaacgtc atgcgaatag tctggatacc
ctagtttgag tttaagttga gagggtttct 1680aaacgatttt tggaaataca ttaactttgg
ttcagacttt ctcgattaat taccattaaa 1740aatcaaaagg tcaatgttgt cgaaaatgta
aaagtatagg cttcaaatag tacaatggta 1800caatcaatgc ccaacgaaac gaacgaagca
tatgaaacaa atatagtggt agatacatgt 1860gatcgtgtat gattagaatt tgctactctc
gaattatttc tgtttagttt ctagagagtt 1920tggttggttt actatttgca gatagatcat
ggagaatgct acttcgacct caatctctgg 1980gaaccatata ctcactaatc atcttcaaac
gatttgtccc aaaacaacgt atattactaa 2040ctgcgtttga tgttttgaat atgtatgata
aggttaaaca tatagagcgt atagattttc 2100aaaagcatgg tagctgttgg tgaaaatgca
tggcttatta gtgtttcttt tagagaagct 2160aacgttgtat atagcattaa tttacatatc
taattttgtc aattgtaatg agactttgga 2220tacaacaaca tccggctgcg aaataattgt
gactttatat gtatttttaa taaattaaat 2280ggtccaacta acacagaaaa cgtacgagta
cactgtctca atatttgttt ctcattggtc 2340cacgtccaac ttggccagcc actgaccact
tcaactacaa tttaaaatga actgacaaat 2400ctccaaccaa acactgaccc gtaataaatt
ctatctatct ttagagactt cgagtccttt 2460cccttttcta ctttcctctt ttggattcat
taattaacta cacaaatcaa tttgaggaaa 2520atcacatgat tctaacacac ttctttcagg
taaagtgtca aaagtttcac agaccatatc 2580aattcaaata ctccatgcga aattaattta
ttaatgctat gaaagtgtgc aaccggtgca 2640cccatgggaa acaatcataa cacaaataat
tggtatacat attttgtaat aaacatggta 2700acaattagaa tggttaaaat gtttaaacag
tgaattgaga taagttttaa ctcgtgatca 2760gaactatttt tatgctcttt tgtttaaaat
ggtcgtattc atgtgttctt atacgtataa 2820agtcacaaaa agacgaccat tttttaaaag
tgcgttcacc tatttgtaca acttgcggta 2880acgattttgt gttttaataa acggtaacct
taaaacgcag accacatttt ccgcggcaca 2940aacgcgctta acgccatctc tataaattaa
aacccaccca atgcccaata ttttctcata 3000t
30011001256DNAArabidopsis thaliana
100aaggtaacac atgtatatat atgtcacata gacattacta gtatatatta tgtacttcta
60tcatatattt atgatattgc agttgcagcg tacacaagtc agctcctttt gacttttaca
120tctcatgaat gcattgccat gacatctaat cttactcgag atttgtgcat gcacattatt
180cacttttgtc ttttgcaatt tttgttattg taaaaaaagg aaaaacaaat gtaaaagaga
240gagagaccag aaaggtctaa ctaaacctaa agagtcaatg aaatgtgttt ctttcttgtg
300ggattaatca attcactctt taacacttct ttataccatt gaagaaatta gatgaaagag
360tcacgagttg cttaccaaat ccctcacaag aattgagaac tgataaacca aattgagaag
420attaaatatc acgtctcctt ttgatctcta ttataattaa tcgaaaataa aaataagagt
480tttcaacaaa acgtgatcat tggtttacga tcacttgcaa agtcaaacct aaaacgtagc
540attagtacac taaccttaaa tactaattat atcatgcaaa ccctaatgtc attacctaac
600tatacatgtg taatgtgttc aacagatctt cttaacccac attagatcaa tattaaacaa
660taaaaagatt cttatatatt ctactactta cttcttctta ttcccatcca tatttttctg
720tgcctttagg ttctcaacta atctcattta atttagctag cacacagaga aacacacacg
780tatataaata atatgataac acacaaaaag actcatatat ataaataatt agagtcatta
840aatgtggatt catcattaaa tgaaacaact cttcttctct gtacaatttc tcttcacacc
900ttcaccaaat tctttgactt caaaaatctt ataaaattta tatatctcca aaaccataaa
960accaaaacga gttttcacaa ataaattact tagttgaaat ttcaaatctc attcaattag
1020ggtacactct ctcaacaatc cacattaatg agggttgctg cttctgatgg ctagcagtaa
1080cagttttatc gcctccactt cttaatgcca tctttttcct cttccctctc cttctctata
1140tatattttct gactctgcaa aaccttaatt catccatctc tcaaacacca tttttggaaa
1200caccatttca tattccttaa acttttccat tttagtatca tttcatattc attgat
12561011219DNASolanum tuberosum 101aagcttttaa acatcgataa ttcatcactt
ttattttttg tactcttctt cttcttcctt 60cctttctttt tttttttttg tgtgaaattt
gatatttttt gtcttaaatg attaatctat 120tgtgtagaaa atagattttc ttgttagtgt
ataaatttta taaaataaat ttaaagacct 180cttaatataa ttttcgctta ggccacgaga
tttgttgagc cgccctgatt atcataaatt 240atttgaagat tttggtctgc aattgtcagc
taatctccaa ctaaataatg tccaacataa 300tttggaccct accaaatatt taacgggcaa
agattaatat aacactatag tatataaaat 360gacattcatg agtgtgaaat tgtatatagt
gttcatgtgc atattttact attttcttgc 420aaatcatatg gttcatatac aataataaca
atggaaaaga caggtgtttg gcctgtaatg 480ggtctctatt gtccagatct tggtggaccc
tacacactat gacgtctgtc aaataatctt 540ggaaaaataa cttgttgcac gactcttcga
gtctaatttt cagtgattta tttaataatg 600actaagtttt atcgctttta taatgacaaa
aaggatttct tattattact atctctgttc 660tatattaatt gaatcgatga gccaattata
tgaaatttta tcaaatattc attttaaatt 720ttgaacgata aaaaaagcct catgagaatt
ttatcaaagt aaaatatgaa aaaaatgatt 780atcaagtaaa aatgaacaaa gagaataata
tgaaggtttt atcaaacatt catcttaaat 840tttgaacgat aaaaaaagct tcgtaaagaa
tattttatca tagtaaaaca tgattatcaa 900gtaaaagtga acaaagggag taatatgaag
atttatcatg tatttaaaag ctcaatagtg 960attataattt gagggactaa ataaatttaa
ggagttgtta atatattccg agaaaataaa 1020atattgttta agtagaaaag ttatggggtg
tataagttaa ataataatat tttgtaaata 1080gggatatgga aatgagtata aatagaaaga
tagcaaggtt tctcgtgaga gttcacaagc 1140caataaagct gatcacactc ccctttgtat
gtccactcaa caacacaact tcttgtgatt 1200cactttcaat tctagatct
12191023430DNAArabidopsis thaliana
102aaaaacaaat ctcatataaa tcatgccgac attacacaaa caaaatcact tatgcagtgt
60ctatgactcg tttatcatgt tgttcttgta ccgttgatat tgtggaaaat tatcttgttt
120ttatttgtgt taaatcagat ctttctcctc ttagtagcat gagaaagagg atcaacccgg
180gacgatattc catccataac gactataatt acagttaatg ttaaactgtt tttatttata
240agacacaatt ataatacatc cattaagatt tttaaagttt tcataacaaa aaaaataaaa
300aatataataa ttagacgcat tttgataata gaaaaactct aaattttcaa taaaagtctc
360aactgccttt acagtctaat atgatcatga gaagggttaa gaatagaata gttgactttt
420aacttaatgg aagatttact ggaaaatgtc tctatctgtt atctagtatt ggacccgtta
480ataatgtgca acgtataatg tagggtacta aatcccaaga aaaatctaga ttttctagat
540aacatattta accagaaaag aaaaaaaatt gtagtggaat agaccagcat ggttcaatct
600ggtcgttact aaaactacca atccaaaacc taccaactat aatacaatat gactacaaat
660acggttttga agattaattc gcggctaacc atcaaaaaat aatgtttgac gatggctcat
720ggttccactc gccaaccata tacagtttat attctctttt gagacctcaa aatcaagtta
780ctagaactcg atgatcttac atgaatataa aatgaaaatc agctccaatg tagtttgaag
840aaggtataaa acaggtttag accaaccgat tccaagactt atgatggccc acaacaacaa
900aaataactaa caaattttta gaccctacac tactcttagc atgcatgtga cagcttaaac
960ttaaatttaa acagttaaca tgaacaaccc cacttgcact aggaagacag caaatagcaa
1020acttaaagca gtttacgatt tctggtcttt taaggcaagg tttagtaacc cagtaattcc
1080ttttgatttg aaatatatga ataaccgaaa ttaaaaaaca aaaaaaaaat cacaaaatac
1140taaaatttga aagccgaaat aattcatttt tcaatagtta taatcatctt ttaaaaagta
1200agttagatca taaaacctaa tataaaactt ttattggcaa aactaattaa tcacgagaaa
1260ataatattaa agttagctca tgttatgttt atattaggaa tctctcattt actttatttt
1320gttttgtttt gtttctataa tgatattagg attcaggaag acaataaaca aaatatcaac
1380aaggaatcat gaaagataca ttcgaacggg ctcagatcga cgcagaaaca aaaagctttg
1440gaatgattta ataatacaaa tttggcggaa aggtttgaag aatttatact tggcattcta
1500tactagcgaa cgaaaacaaa agtttagtca aatatttagg attaccaaaa aatcaatatt
1560cgtatacaaa tgtgttttat tataaaccat gatttttcat aactttattc ttagttgttt
1620caatttgttt tctctcattt ctcgttccta ttgtagaaaa aattcctgat tcttttttct
1680tttcttttgt tttcccatgt caattataac cgcattagaa aaggttagca catgttcatt
1740tgcacagaat gtagtcaatg tacaattcat ctataattta tcaccttttt gttaaaaata
1800tttatagcca tgttatatat attttatcat gttgtgtttt aaactctaga tttgttcttt
1860cacagtttat gggatttatc atttcatata tttcctccta tgagtaagat gtcattttta
1920aattcttaat taagaattta gaaatcaagt ttattatttt gttaaaattt ataaattacg
1980tatttttcct ttattagtga gattaagata taaagagtct agcacactag agaaatgaaa
2040acatacatat atctagaaag ggtttataag aaattagtaa atggaatcag aacatgaagg
2100agaagggagg ggccaccata ccgacgggaa tgagaagtat gggacggaca agaaatcaag
2160tgtacacaca tgaaaaaacc caccacgtga agatctccta ttggtgtgtc ttctcctaag
2220tactaagtat aaaccaaagt caaaacacaa caaagacata ttgcttgcga gccagcgagt
2280gtgtccgagt gagaagagag tttgggactt cactgtttag agagcctcat tcgctaccta
2340gattcggttt atccacgtgg agagttcacg tagcttctat aataataata cattatataa
2400gctgtttcgg taaaataatc ttagagtttc acttcccatg caaaaagatt aaatggacta
2460ctatattttc tatatgagtg ttttggaaat aatcttccaa gtttatggct atttagtaaa
2520actcaatcca aaatgagaac aataactttt tttttttttt ttttgcaatg agtacaataa
2580acttatgatt ggttttgggg actagaaaat aaaatagaga catatacttt aagaatagtt
2640gatgagaaaa taaaactttt ttcgaagaaa gaaaaaaaaa tatactctgg ccccttacat
2700attctgtcaa atgccatttt attttcttta ttttgcttgg tccatttctg gattcctgaa
2760atgaacacac cataattttc atgcccattt ttgtcctccc attaacactt accaaacacc
2820aaacgtacct tccttccttt atatgtgtct ctatgtctat atatcactcc caccttcaaa
2880tgctaggaaa gtggacatca caccatagtc tgatagtcat gaatctcatc aagtagaaac
2940ctccaaagaa acagattgcg tttcctcttc ttcttcctcc tctttcgaat cttatatttt
3000cgtgtcaaaa aagaaacaaa atatgatgat atctaccgca aaagctcaat gtttgccact
3060taaattctgt ctcatctctc aagatttttg atcatctcac acttactccg ttccgcattt
3120tacgcctttt cttcaccatt gttgcttctc gaatcgaaac catattggtt ttgcagcatt
3180aatggtgcac tgagtcttct taaagccgtt aacctttccc tgccaaattc tcaaccaaaa
3240agaaaaaaag ataacaaatg ggtcacaaca agttgccata aaggtcccaa cagaagaaga
3300aaagagaagt ttccccgtgt tgctttgtca caagcttcat cactatttat aactaccttg
3360ccaagaagaa agcctaaaaa ggtataacag tttctttgtc tttagagatc atcaaaaaga
3420tgtcgatctc
34301032630DNAArabidopsis thaliana 103ttggatccac caattattaa caaggagaag
tgttatagga acatgagaaa gcaagaagga 60agctaagatc atcttttcat atgcccaaaa
ttccctagtt agtcttgttt agttcatatt 120ttcttgtacc taattgcttt atttcatgta
agcaatatcg catgtacgta tacgatgtat 180ttttatataa ccctagttct taaatatttg
taatgcttat taaaaatcac aggttttcat 240cttaaaacaa attggcaata agtaaaaaaa
tttaatgttg aatttatcaa acttccaatg 300tgggacaatt tccctatcta ataagtcttt
gagaatttgt gagaaacggt ctaacaagaa 360caatccaaac ttaatatatt ggaaatacca
taacaagacg gatcacttat acgaaccgca 420gaggtgagaa acccctttat ttggaaaagt
gtctagtggg tcaatctggc tctgatacca 480tattagaaat tataggcttt caactcaaaa
ccaattgaca atgagaggag aatattagtg 540tagaatttac tacaagcatt atataatatt
aatatatatc aaattacaac tttaaaatat 600aaagtatcat attttgtttt attcctaata
tgatctaatt taggaaataa aaactctaat 660acatgtatag tacgtacact gatatgtgcc
ctcttgggag gaaaattgcg gaaccgcatt 720tttgcgggaa aaatattgac tcacgttttt
gtggaaacta tacggaatca catatttttg 780cgagaaaaat tcattctctt atttttagct
ggaaattatg aaccgtattg ttagcaaaga 840actatagaat tatgttttga cgctaaaaag
cgaactcgca ttttttgcgg aaaaatggtg 900agatcgtgta tttgacagaa attttgattt
tgagagagaa tgagagatcg atgatgaaag 960atgtgagaaa tttgtgtttg cttctgttta
ctaaaattgg tttaaacatt ttattcagtt 1020atatttggtt tacacatttg gttattttgt
caatttggtt atgggtaatc cactaatcta 1080tttcaaccaa ttattccaac ccaaatcatt
acactcatta atccattgat tcattaatta 1140tcaaatttcc aacaactcat tagatccatg
agttggattg gtttgggttg agttgaaaaa 1200ttaacttcaa ttcattttga catacctata
atatggtata tcatacgaac cacctaaaat 1260atgtgatgca tgcacttatt aaattgcatt
ttacaccatg tttttgttta agtacgtcaa 1320cattgaatta gcacaatgta tacgtacaaa
aaagactact ttcatggtaa tttaatattc 1380taactcaaca agatatcata gaatgcaata
agaagttgtc attgcaatga ggtactatca 1440aaattctaaa agtctcaatt gtgtaatttt
ccctaagaaa tagggaaaat accatttaaa 1500tatcaaacat ttacaaaatg ctaaacaatg
ttcaaatttt ctttctatac ttcttaagtt 1560ttaaatacga aatttctttc acagtaccaa
attaaaactt tgttaatatc atcggatatt 1620tctaaaaaag gaaataattt acatgatcct
caatccccga aaaaaagtat ggaagataac 1680agttctgcat atgataaagc acctccaatt
ttaataacga ttaaaaaaaa gcaaaagggt 1740ggttggttta tatttattgt cttttggata
gagagtctct agtttcatgt gataaagtat 1800cgatcgccca aaatccattc atacgagata
tgctgtgctg tgaggtgtga gcggtaaggc 1860tcgtgatgaa tggcaaacga ttcatgtaac
tattttcttt tattatctta acaaagtcgt 1920tgttactcta aatttaacac ttgatcatat
ttctaatatg tatattctga ataatggcat 1980ggcgtcaatt ctaacttatg agactttaaa
cttataatgt tcactttctc ggtagcaaaa 2040acgcttctct ttgcccctta agtagaagga
caagaaatct attttttgtt tcattgctcc 2100cgtttcctca catctgcact tgctaagcca
accttggctt acttatgacc acctcgagaa 2160gaacctattt gaaaacagtc taatgccaca
aaccaatctt gatgcatgga cttctagttt 2220tgcgagaacg tttaatacat attatactgt
atgactgttg accaatcaat tgaaaatgtg 2280acttagtttg tgctgcaagt cgtgttccgt
atgaggataa catgagataa cgatagtaat 2340taacagttca caactgagta aaagaatgca
acacattaaa atgatcaatt gaaaagcaaa 2400gcagtccgaa aagtgtaaac aaacatttaa
aaataaaaaa ttgttacatt caggtagagt 2460aagcaatccc acgggattca tgtcacctta
acctcccaaa atatcatgac gctaagacaa 2520ggtaatgtat acgtcactct tctctaacaa
aggctatata aaccctagat tggcactagg 2580cttccatgca acacaaaaac atacacacta
ctaataaacc atgaagcttc 26301042001DNAArabidopsis thaliana
104tcaatcttat gagttgattt cggaattgtt ttggcttcat cttttttttt tttttatgtt
60tgttttgttg ttgtttaaac tgtttcgatg actctttgta tatggtggga gttttctttt
120gattatgtaa tgtttttttt ataattactt ttcttctttt ggattaatga ttattcctga
180ggtggagact ttactatata tagagagaat gagttggaag atggaacttg ggaacaaatc
240gtcgtggaat acgaggtgta ttagactttc agttgtgcaa agggacaaaa caacatgatc
300tgttaaaatt tctttgttta tacaccacaa attaatgcat acacacacag tatacatgtg
360taacgttttt gatgttcccg tgaaaataga tgtgtattta gagcctagat ggatagatta
420tgattggatg aaacatttgt aatattttga caaaagttta ccatttacgt taaatgtaat
480agaatgaaca ttcctctaaa aaaaaactcc ctaagcaaaa aattactata aggatgaatg
540gtatatcgct tagtcacgtt ttcaaacgct atttttttac attcaaaata tttatcaaat
600ttttttcgat ttcacgtttt cgaaaagtaa ttcggctgta tactactagg ctactagcat
660gtcaatatac ctagaaaaaa acatgattaa tccactagaa ctatattttc atccattaaa
720ctaatttatt cagttcaatt cattccacta ataattattc agttattccg tcatttcact
780ataattataa gttatagaca agtttttttt actaacatgt tatagctatc cttcaaaaat
840ggtacttagc aataacatct tatagctatt ctgaacatcc ataatgtttt gtcatccaat
900tttgttcttc aaaataaaca agttaaaatt aatttctcgt cttcaattct aaataattgg
960ctccactgat gaaagcccat aacccatatg gaaaaacgtg catttcagca tttgtgtaga
1020tagcgatatt ttgttcgact acacagaaag ttttttttct ttttgactac caaagaaatt
1080tgattcatag tatcttcgag ttaggtacca aaacatgtaa aacagtctcg tcttaaaccg
1140tccaatacga cttcatgacc gacaaaatta aatcacacgg ccagttttga cgcgtgagac
1200aacaacctac tattgaggat ttggctaaac actcgccaca atgcttctat tgtattgaca
1260atatttgtgt ttatttcatt tgttattaat aaagacaaat atcataagta ataattgact
1320actactaatc cataatttaa aatgttattt tttttgaacc aaggactaca taactaacaa
1380cttgtgacta aagattacaa aacatgtttt agtcgttatt tttaatcatt atagtttata
1440gtaatcgcta ttattcgttg ttattgctaa ctatgaaatg aacatggttg tacattttgt
1500cccatcacta ccaccaaaat ttttggattt tgttgaaaaa agggagacta accatggttt
1560acaaagaatc tatgaaatac taacgaaact atacacaatt atcgacttta ttattaagag
1620acatcgtcta caatatatga aggtcagcgt gtaacatatt tttgtttttg gggcgttttt
1680agtattttag acaaatatag ggatttatcg taatttcgat aatcttgtgg tttttttagt
1740tataagtctc ctcttgtcat attcacgagc cgtataaaaa agttgtttct ttcttctctg
1800tttttttgct ttcgtcttca agagagagag agagagagat acaaagagag aaatttggtt
1860gtttgttgac ggaagcttct tcggtctctc ttctccgtct tacgattgtc aacgcgtggt
1920tccatcttca attttgtttc tattttagca gaagtttctc gagcttcaaa tactgtttca
1980gatcaatcaa tcagtcaatc a
20011052519DNAArabidopsis thaliana 105ttttggggat ttcaaaggac tctcttatcg
acgacttttt ttttttgttt tgttctcgtc 60tatatttgga acatgtgata tagttcaagt
atgaagaaga tatgtgaaga aatttatagg 120caaaaattat aaatgtattg gtttgaagat
tcaaaaaagt aattatgttg ttacttgtaa 180ttttggtgca tctcacgtgt gactagacct
tgagtggaag tatttggttt tttttttttt 240tcttatgata tttgtttttt tagtagaaaa
tttaaaaacc attagatttt gggtgctata 300attagaaacc atctatgtct ttgaattgtt
catgtttaca ataaagtata gaaagtaaaa 360atctactcaa aaacaatttg tggttgctat
aactcattct tatcagtttt atttcaagca 420aaactaataa cataaattta gtattgagaa
gaaaaaaaga aacaactcgt ctcacagacg 480gaaattgcaa actaaaataa gagtggctta
tcaccgaagc aggaatcatg tgagtcaaac 540gtgggacgcg agagatgggc gttgacaaat
gctgcgtcgg cgtggatggg ttttacaagt 600gcaaccaaat tgacacgtat ctatgacttc
tcatatattc caaagtattt cgatataagt 660tggcaatatt ttcattaaat agagctgttg
atttttccaa gaagtggtat ctcaattctg 720cttggtattt acgactttcc taacttttta
tctagtttag attgttcgtc aagtttgaaa 780taaaactgtt gataatagct atgactatca
tatacagtat gaaaaaatta agagttagat 840tgtggcaaag agacatctga aattgtagaa
ataaaggatc taaagcagtg gactgcctcg 900aagatgctat aaattttgta atcacacttt
atttcgacca tttcatggtt aagtaattat 960gtatgtttgt ggtttaagat taatataatg
atcacacggt tttctattag atgttttgga 1020gcgttagtta cttttgtgca tattatctta
aggatctctc aaatccgatt gacgcgataa 1080aaaggtcacc aaaagttata gactcgaaat
ccatattttc tgacctacct aatggtcgaa 1140atattagtcc aagacacata tccaatggaa
aaaaacttat ttaaattttt taagagaaac 1200tacgtcaatc atattacttt ttaaaagttt
agatagtggg agaattcaca agaatactga 1260ttacaaaaaa aaaaaaaaaa aaaaaaaaaa
aactgaaaat cacgaggcga tcgttatttc 1320atttcccttt ctatgaaata gatttggact
tttcatgaag gcaagccaaa agtgccaaac 1380aagtggtcct ccaaattttg aggaacagtt
agttcaaaac atagcaagtt tggtctcacc 1440catctttttt tacattacag ctcaaagata
gaacattaaa aaaaaaaaga aaaaaaaaga 1500tttatagaga agattcatga atctagctag
actattgtac cttaaattgc agctcaaagt 1560catattttgc atgttgatca gatatgtcat
aagagttatg tcaaatacac ttgttgtatc 1620tctttgtaga aaataagatt gctaatccta
ataattttgt tgacaaaaaa aagaaagaaa 1680agaatacatc ctcaatttat aaagacacta
tagctttgac attgacgact cacacgccca 1740aataaggtcg gcagagtgaa attgtctgtg
aaatcatggt gttttaagtt gtactggaac 1800ttgtgtatga aacagtctat agcaataata
atgaaaaaga aacaacacat tttgctccat 1860agcttttgta tttgtgtaaa tcggaaagaa
aagtggttta ttattcatgg tcgaaaaaat 1920cagaaaaatg ggtcgattag agaaaaaagt
aattttcagt ggctacagta taagtacagc 1980gaactgttct aggtagagag tcccattata
caacaacaac tcattataaa atttgacttc 2040agtaacgact gattgagaat atgttaatgt
acactaaact attgacattg atgtaattgt 2100atatttttgt acaattacgt tagtaatatg
gcgattgcac ctgattggtg aagaatctat 2160attctcttcc agttacccta cactagaatt
tttcaatgaa gttatcactt gacataatca 2220atttaaaaat ttgatttcga gacttcgacg
ataaattttg ttgggccact acaaaaggtt 2280ttactgagtg ctgactactt attataatag
gcccaaacaa aacatatatg ttgggccact 2340acattattag ccaaaagatt ttactgtaac
ttattatgag cccatacgga gcatttcaca 2400gggaaaattt aactaaacgc gaaagtggcg
ttacgattat tagaatgatt tcgtaataaa 2460cagaggatta gttaaatcac gtttcgatta
ctgtatatga ttaaaaatta aagttgaag 25191063497DNAArtificial sequenceA
construct sequence consisting KST1 promoter, AtHXK1 cDNA and a
terminator 106gtcgactaga aaatgaaatg aaaaacacct atctgttttt tcactcaaat
tccatccttg 60caataaaatg cttattctta aaatttctat cttggtggag atcccaccac
cattaccatt 120ttccccaaaa atcttacaac attatttcca ttttctttct cttaattctc
tcaacaaatt 180ccccttgcac ctgaattatt atcaaagaaa atcatgtttg ccacttcaac
aactttataa 240ctcatatctc gcctcgtttg ttgtttattg tgtttcaata ttgctatgtt
ttcttctatt 300ttgtacttgc atttgctcac tcgagctttt ggtaacaatc tctctacttc
tactagatct 360gcgtacagtc taccttctcc agaccccact tgtgggaaga tactatagaa
gtaggcaagt 420agcaatgtca cgttcttaaa gctaaatgct ttttcaaaag aatcacaata
aagaaacact 480tgacccgtgt atcaccccaa ctacttcttc atctacatcc tctatatata
aacacgctaa 540aaataactag ttagtatttt taaatattac acattgcctt tccaagaaac
tcgaaaaaaa 600aaaaaaaaaa aaaaacccac atcaacaaaa aagaagcagc aatatataat
actgcagacg 660cgtctcgagg aattcggtac cccgggttcg aaatcgataa gcttggatcc
cgccagtgtg 720agtaatttag gtttctctaa tttctctcaa ttcactccaa aattttgatt
atttcttctt 780tctggcttgt caattttagt catttgtaat ccttgctttt gcgatcggaa
tcgtaaaaat 840ccgatctttc ttttagattc gttttgtttt tgattccaaa tcggaaaaat
gggtaaagta 900gctgttggag cgactgttgt ttgcacggcg gcggtttgtg cggtggctgt
tttggttgtt 960cgacgacgga tgcagagctc agggaagtgg ggacgtgttt tggctatcct
caaggccttt 1020gaagaggatt gtgcgactcc gatctcgaaa ctgagacaag tggctgatgc
tatgaccgtt 1080gagatgcatg ctggtcttgc atccgacggt ggtagcaaac tcaagatgct
tatcagctac 1140gttgataatc ttccttccgg ggatgaaaag ggtctctttt atgcattgga
cctagggggg 1200acaaacttcc gtgtcatgcg tgtgcttctt ggcgggaagc aagagcgtgt
tgttaaacaa 1260gaattcgaag aagtttcgat tcctcctcat ttgatgactg gtggttcaga
tgagttgttc 1320aattttatag ctgaagctct tgcgaagttt gtcgctacag aatgcgaaga
ctttcatctt 1380ccagaaggta gacagaggga attaggtttc actttctcgt ttcctgttaa
gcagacttct 1440ctgtcctctg gtagtctcat caaatggaca aaaggctttt ccatcgaaga
agcagttgga 1500caagatgttg ttggagcact taataaggct ctggaaagag ttggtcttga
catgcgaatc 1560gcagcacttg ttaatgatac cgttggaaca ctagccggtg gtagatacta
taacccggat 1620gttgttgctg ctgttatttt aggcactggg acaaacgcag cctatgttga
gcgtgcaacc 1680gcgatcccta aatggcatgg tctgcttcca aaatcaggag aaatggttat
aaacatggaa 1740tggggaaact tcaggtcatc acatcttcca ttaaccgagt ttgatcacac
gctggatttc 1800gagagtctga atccaggcga acagattctt gagaaaatca tttccggtat
gtacttggga 1860gagattttgc gaagagttct tctaaagatg gctgaagatg ctgctttctt
tggcgataca 1920gtcccatcta agctgagaat accattcatc attaggactc ctcacatgtc
ggctatgcac 1980aacgacactt ctccagactt gaagattgtt gggagcaaga ttaaggatat
attggaggtc 2040cctacaactt ctctgaaaat gagaaaagtt gtgatcagtc tctgcaacat
catagcaacc 2100cgaggagctc gtctctctgc tgctggaatc tatggtattc tgaagaaact
gggaagagat 2160actactaaag acgaggaggt gcagaaatcg gttatagcca tggatggtgg
attgtttgag 2220cattacactc agtttagtga gtgtatggag agctcactaa aagagttgct
tggagatgaa 2280gcttcaggaa gcgttgaagt cactcactcc aatgatggat caggcattgg
agctgcgctt 2340cttgctgctt ctcactctct ctaccttgaa gactcttaaa acctacccaa
agagcgccat 2400ttttcggtaa tttactgaaa gcttttcgct atcagaaaac gcctaagcca
agttctaagg 2460cgtcataaaa gaaagcattc catgttttta ctcttcccca agactttctt
tgtagcaaat 2520aagtttcctt gggagaaata tttgttttca tgttcttcaa aaataaaaga
ctcagttctt 2580cagattctgg gattttatta taaccagata tgttgtaaaa actacaaatt
caaagctcac 2640ttcactggag ttctgagtat ataaagattt catttttcct aaaaaaaaaa
ctaaattact 2700cacactagcg ggatccatgc tagagtcctg ctttaatgag atatgcgaga
cgcctatgat 2760cgcatgatat ttgctttcaa ttctgttgtg cacgttgtaa aaaacctgag
catgtgtagc 2820tcagatcctt accgccggtt tcggttcatt ctaatgaata tatcacccgt
tactatcgta 2880tttttatgaa taatattctc cgttcaattt actgattgta ccctactact
tatatgtaca 2940atattaaaat gaaaacaata tattgtgctg aataggttta tagcgacatc
tatgatagag 3000cgccacaata acaaacaatt gcgttttatt attacaaatc caattttaaa
aaaagcggca 3060gaaccggtca aacctaaaag actgattaca taaatcttat tcaaatttca
aaaggcccca 3120ggggctagta tctacgacac accgagcggc gaactaataa cgttcactga
agggaactcc 3180ggttccccgc cggcgcgcat gggtgagatt ccttgaagtt gagtattggc
cgtccgctct 3240accgaaagtt acgggcacca ttcaacccgg tccagcacgg cggccgggta
accgacttgc 3300tgccccgaga attatgcagc atttttttgg tgtatgtggg ccccaaatga
agtgcaggtc 3360aaaccttgac agtgacgaca aatcgttggg cgggtccagg gcgaattttg
cgacaacatg 3420tcgaggctca gcaggacctg caggcatgca agctagctta ctagtgatgc
atattctata 3480gtgtcaccta aatctgc
34971072280DNAArtificial sequenceA construct sequence
consisting KST1 promoter, GFP cDNA and a terminator 107attcctgcag
cccgggggat ccactagttc tagagcggcc gcatgcatat gtcgactaga 60aaatgaaatg
aaaaacacct atctgttttt tcactcaaat tccatccttg caataaaatg 120cttattctta
aaatttctat cttggtggag atcccaccac cattaccatt ttccccaaaa 180atcttacaac
attatttcca ttttctttct cttaattctc tcaacaaatt ccccttgcac 240ctgaattatt
atcaaagaaa atcatgtttg ccacttcaac aactttataa ctcatatctc 300gcctcgtttg
ttgtttattg tgtttcaata ttgctatgtt ttcttctatt ttgtacttgc 360atttgctcac
tcgagctttt ggtaacaatc tctctacttc tactagatct gcgtacagtc 420taccttctcc
agaccccact tgtgggaaga tactatagaa gtaggcaagt agcaatgtca 480cgttcttaaa
gctaaatgct ttttcaaaag aatcacaata aagaaacact tgacccgtgt 540atcaccccaa
ctacttcttc atctacatcc tctatatata aacacgctaa aaataactag 600ttagtatttt
taaatattac acattgcctt tccaagaaac tcgaaaaaaa aaaaaaaaaa 660aaaaacccac
atcaacaaaa aagaagcagc aatatataat actgcagacg cgtctcgagg 720aattcggtac
cccgggttcg aaatcgataa gcttggatcc atggtgagca agggcgagga 780gctgttcacc
ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa 840gttcagcgtg
tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt 900catctgcacc
accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta 960cggcgtgcag
tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc 1020cgccatgccc
gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta 1080caagacccgc
gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa 1140gggcatcgac
ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa 1200caggccacaa
cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca 1260agatccgcca
caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca 1320cccccatcgg
cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg 1380ccctgagcaa
agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg 1440ccgccgggat
cactcacggc atggacgagc tgtacaagta atctagagtc ctgctttaat 1500gagatatgcg
agacgcctat gatcgcatga tatttgcttt caattctgtt gtgcacgttg 1560taaaaaacct
gagcatgtgt agctcagatc cttaccgccg gtttcggttc attctaatga 1620atatatcacc
cgttactatc gtatttttat gaataatatt ctccgttcaa tttactgatt 1680gtaccctact
acttatatgt acaatattaa aatgaaaaca atatattgtg ctgaataggt 1740ttatagcgac
atctatgata gagcgccaca ataacaaaca attgcgtttt attattacaa 1800atccaatttt
aaaaaaagcg gcagaaccgg tcaaacctaa aagactgatt acataaatct 1860tattcaaatt
tcaaaaggcc ccaggggcta gtatctacga cacaccgagc ggcgaactaa 1920taacgttcac
tgaagggaac tccggttccc cgccggcgcg catgggtgag attccttgaa 1980gttgagtatt
ggccgtccgc tctaccgaaa gttacgggca ccattcaacc cggtccagca 2040cggcggccgg
gtaaccgact tgctgccccg agaattatgc agcatttttt tggtgtatgt 2100gggccccaaa
tgaagtgcag gtcaaacctt gacagtgacg acaaatcgtt gggcgggtcc 2160agggcgaatt
ttgcgacaac atgtcgaggc tcagcaggac ctgcaggcat gcaagctagc 2220ttactagtga
tgcatattct atagtgtcac ctaaatctgc ggccgccacc gcggtggagc
2280108686DNAArtificial sequenceKST1 promoter 108gtcgactaga aaatgaaatg
aaaaacacct atctgttttt tcactcaaat tccatccttg 60caataaaatg cttattctta
aaatttctat cttggtggag atcccaccac cattaccatt 120ttccccaaaa atcttacaac
attatttcca ttttctttct cttaattctc tcaacaaatt 180ccccttgcac ctgaattatt
atcaaagaaa atcatgtttg ccacttcaac aactttataa 240ctcatatctc gcctcgtttg
ttgtttattg tgtttcaata ttgctatgtt ttcttctatt 300ttgtacttgc atttgctcac
tcgagctttt ggtaacaatc tctctacttc tactagatct 360gcgtacagtc taccttctcc
agaccccact tgtgggaaga tactatagaa gtaggcaagt 420agcaatgtca cgttcttaaa
gctaaatgct ttttcaaaag aatcacaata aagaaacact 480tgacccgtgt atcaccccaa
ctacttcttc atctacatcc tctatatata aacacgctaa 540aaataactag ttagtatttt
taaatattac acattgcctt tccaagaaac tcgaaaaaaa 600aaaaaaaaaa aaaaacccac
atcaacaaaa aagaagcagc aatatataat actgcagacg 660cgtctcgagg aattcggtac
cccggg 686
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