Patent application title: PERMISSIVE CELLS AND USES THEREOF
Inventors:
Peter Delputte (Kortrijk, BE)
Hans Nauwynck (Zomergem, BE)
Hans Nauwynck (Zomergem, BE)
Hanne Van Gorp (Gent, BE)
Assignees:
UNIVERSITEIT GENT
IPC8 Class: AC12N702FI
USPC Class:
4242041
Class name: Drug, bio-affecting and body treating compositions antigen, epitope, or other immunospecific immunoeffector (e.g., immunospecific vaccine, immunospecific stimulator of cell-mediated immunity, immunospecific tolerogen, immunospecific immunosuppressor, etc.) virus or component thereof
Publication date: 2014-07-03
Patent application number: 20140186395
Abstract:
Described are methods for determining the permissiveness of a cell for a
virus that is a member of the family Arteriviridae or Coronaviridae or
Asfarviridae, in particular, for Porcine Reproductive and Respiratory
Syndrome Virus (PRRSV). Further described are methods and compositions
related to the generation of host cells permissive for a virus that is a
member of the family Arteriviridae or Coronaviridae or Asfarviridae, in
particular, for PRRSV. Methods of utilzing the cells thus identified or
thus generated, in preparing a culture of a virus that is a member of the
family Arteriviridae or Coronaviridae or Asfarviridae, as well as the use
of the virus for the purpose of vaccine production or diagnosis, are also
described.Claims:
1.-30. (canceled)
31. A method for generating a cell that is permissive for Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), the method comprising: treating a cell to express CD 163 and sialoadhesin proteins, thereby generating the cell that is permissive for PRRSV.
32. The method according to claim 31, wherein treating the cell comprises introducing into the cell an exogenous nucleic acid encoding CD 163 protein.
33. The method according to claim 32, wherein the CD163 protein encoded by the exogenous nucleic acid is at least 70% identical to porcine CD163 encoded by SEQ ID NO:1.
34. The method according to claim 32, wherein the CD163 protein encoded by the exogenous nucleic acid includes at least one Scavenger Receptor Cysteine Rich (SRCR) domain.
35. The method according to claim 31, wherein treating the cell comprises introducing into the cell an exogenous nucleic acid encoding sialoadhesin protein.
36. The method according to claim 35, wherein the sialoadhesin protein encoded by the exogenous nucleic acid is at least 70% identical to porcine sialoadhesin encoded by SEQ ID NO:9.
37. The method according to claim 36, wherein the sialoadhesin protein encoded by the exogenous nucleic acid includes at least the N-terminal domain of porcine sialoadhesin.
38. The method according to claim 31, wherein treating the cell comprises introducing into the cell exogenous nucleic acid(s) encoding CD163 and sialoadhesin proteins.
39. The method according to claim 31, wherein treating the cell to express CD163 and sialoadhesin proteins comprises chemical treatment.
40. The method according to claim 39, wherein the chemical treatment results in the expression of sialoadhesin protein in the cell.
41. The method according to claim 31, wherein the cell is selected from the group consisting of an insect cell, a yeast cell, a porcine kidney (PK) cell, a feline kidney (FK) cell, a swine testicular (ST) cell, an African green monkey kidney cell, a MA-104 cell, a MARC-145 cell, a VERO cell, a COS cell, a Chinese hamster ovary (CHO) cell, a baby hamster kidney cell, a human 293 cell, a murine 3T3 fibroblast, and a plant-cell based production platform.
42. The method according to claim 31, wherein the cell is not an alveolar macrophage.
43. A cell that is permissive for Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), generated by the method according to claim 31.
44. The cell of claim 33, wherein the cell is selected from the group consisting of an insect cell, a yeast cell, a porcine kidney (PK) cell, a feline kidney (FK) cell, a swine testicular (ST) cell, an African green monkey kidney cell, a MA-104 cell, a MARC-145 cell, a VERO cell, a COS cell, a Chinese hamster ovary (CHO) cell, a baby hamster kidney cell, a human 293 cell, a murine 3T3 fibroblast, and a plant-cell based production platform.
45. The cell of claim 44, wherein the cell is a PK-15 cell, a CHO cell, a BHK-21 cell, a MARC-145 cell, or a Hek293t cell.
46. A method for producing Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), the method comprising: infecting the cell of claim 43 with PRRSV.
47. The method according to claim 46, wherein the method further comprises harvesting PRRSV from the cell.
48. The method according to claim 47, wherein the method further comprises inactivating the harvested PRRSV.
49. A method to determine infection of a subject with Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), the method comprising: contacting the cell of claim 43 with a sample taken from the subject; culturing the cell under conditions suitable for replication of PRRSV in the cell; and determining the presence in the cell culture of PRRSV, wherein the presence of PRRSV indicates that the subject is infected with PRRSV.
50. The method according to claim 49, wherein the method further comprises treating the subject for PRRSV infection.
Description:
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of co-pending U.S. patent application Ser. No. 12/452,675, filed Jan. 13, 2010, U.S. patent Ser. No. ______, which application is a national phase entry under 35 U.S.C. §371 of International Patent Application PCT/EP2008/006045, filed Jul. 23, 2008, designating the United States of America and published in English as International Patent Publication WO 2009/024239 A2 on Feb. 26, 2009, which claims the benefit under Article 8 of the Patent Cooperation Treaty and under 35 U.S.C. §119(e) to Great Britain Patent Application Serial No. 0811278.1 filed Jun. 19, 2008, and to European Patent Application Serial No. 07014842.4 filed Jul. 27, 2007, the disclosure of each of which is hereby incorporated herein by this reference in its entirety.
TECHNICAL FIELD
[0002] The disclosure relates generally to the field of virology. More particularly, it relates to methods for determining the permissiveness of a cell for a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular, for Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). Further provided are methods and compositions related to the generation of host cells permissive for a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular for PRRSV. Methods of utilizing the cells thus identified or thus generated, in preparing a culture of a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, as well as the use of the virus for the purpose of vaccine production or diagnosis, are also provided herein.
BACKGROUND
[0003] A "mystery swine disease" appeared in the 1980s, and is present ever since in pig industry causing important economical damage worldwide (Neumann et al., 2005). The causative agent, designated PRRSV, was first isolated in the Netherlands in 1991 and shortly after in the USA. It is a small enveloped positive-stranded RNA virus that is classified in the order Nidovirales, family Arteriviridae, genus Arterivirus together with equine arteritis virus, lactate dehydrogenase-elevating virus and simian hemorrhagic fever virus based on similar morphology, genomic organization, replication strategy and protein composition. In addition, they share a very narrow host tropism and a marked tropism for cells of the monocyte-macrophage lineage (Plagemann and Moennig, 1992). More specifically, in vivo PRRSV infects subpopulations of well-differentiated macrophages, with alveolar macrophages being the primary target cells, although in infected boars also testicular germ cells have been shown to allow PRRSV replication (Sur et al., 1997). In vitro, PRRSV replicates in primary cultures of alveolar macrophages and peripheral blood monocytes (PBMC), although PBMCs need treatments to improve infection (Delputte et al., 2007). Furthermore, African green monkey kidney cells and derivates thereof (Marc-145 and CL2621) have been shown to sustain PRRSV infection, although they are not from porcine origin and do not belong to the monocyte-macrophage lineage (Kim et al., 1993; Mengeling et al., 1995). Notwithstanding this very restricted cell tropism of PRRSV, the virus is able to replicate in several non-permissive cell-lines upon transfection of its viral RNA, indicating that cell tropism is determined by the presence or absence of specific receptors on the cell surface or other proteins involved in virus entry (Kreutz, 1998; Meulenberg et al., 1998).
[0004] So far, two PRRSV receptors were identified on macrophages, namely heparan sulphate (Delputte et al., 2002) and sialoadhesin (Vanderheijden et al., 2003; Wissink et al., 2003). In addition, Wissink et al. (2003) found a 150 kDa protein doublet to be involved in PRRSV infection of macrophages, however the identity of the N-glycosylated proteins is still unknown. In the current model for PRRSV infection of macrophages, PRRSV first binds to heparan sulphate most likely leading towards virus concentration. However, this first binding is rather unstable and is followed by binding to sialoadhesin and subsequent internalization (Delputte et al., 2005). Upon internalization, the virus is transported towards endosomes were a drop in pH is required for proper virus replication (Kreutz and Ackermann, 1996; Nauwynck et al., 1999). Despite this elegant research, the model is still incomplete. Transient expression of sialoadhesin in non-permissive PK-15 cells results in binding and internalization of the virus, but fusion and uncoating of the virus particles was not observed (Vanderheijden et al., 2003), indicating that other proteins are needed for virus disassembly, essential for virus replication.
[0005] PRRSV infection of Marc-145 cells makes use of a heparin-like molecule on the surface of Marc-145 cells (Jusa et al., 1997), resembling the initial step of PRRSV infection of macrophages. However, since sialoadhesin is absent from Marc-145 cells, virus entry will differ between the two cell-types. In Marc-145 cells, the intermediate filament vimentin has been described to bind to the PRRSV nucleocapsid protein and it has been suggested to interact with other cytoskeletal filaments to mediate transport of the virus in the cytosol (Kim et al., 2006). Recently, CD151 was found to interact specifically with PRRSV 3' untranslated region (UTR) RNA (Shanmukhappa et al., 2007). CD151 was proposed to be possibly involved in fusion between the viral envelope and the endosome or to relocalize the ribonucleoprotein complexes to promote viral replication. Still, further research is needed to elucidate their precise molecular modes of action during PRRSV infection.
[0006] Recently, the scavenger receptor CD163 has been described to play a role in PRRSV infection of Marc-145 cells and to make some non-permissive cells somewhat susceptible to PRRSV upon expression (Calvert et al., 2007), where others remain unproductive upon infection, despite expression of CD163 (Calvert et al., 2007). Although the CD163 gene was originally isolated from macrophages, thus far no role for CD163 in PRRSV infection of its primary target cells has been shown. Also, the mechanism by which CD163 confers partial susceptibility of selected cell types to PRRSV infection was not elucidated.
DISCLOSURE
[0007] We demonstrated that both sialoadhesin and CD163 are involved in PRRSV infection of macrophages. In addition, expression of recombinant forms of both CD163 and sialoadhesin in non-permissive cells renders all of them susceptible to PRRSV infection resulting in the production and release of infectious progeny virus. In contrast, when only CD163 is present, infection is clearly less efficient, and even absent in some cell types. In addition, viral adaptation that leads to antigenic differences in viral strains grown in cells only expressing CD163 when compared to the wild-type viruses, has been reported.
[0008] Based on detailed analysis of the kinetics of PRRSV infection, both in primary macrophages and in cells expressing sialoadhesin and CD 163, a role for CD 163 in virus fusion and uncoating is proposed. Compared to the above mentioned systems, i.e., cells solely expressing CD163 or sialoadhesin, it has been found that the combination of CD163 and sialoadhesin expression in one cell provides permissive cells that are highly efficient to sustain viral replication, and which closely mimic the entry of the virus in the natural host, i.e., the known subpopulations of well-differentiated macrophages, in particular alveolar macrophages being the primary target cells of the virus. Such mimicry of the entry of the natural target cells will certainly reduce or avoid virus adaptation in cell culture and the associated genetic and antigenic changes that might result in viruses with altered epitopes. Such modified epitopes can have tremendous effects on the antigenicity of vaccine viruses produced on given cells, resulting in loss of induction of important neutralizing antibodies. Clearly, avoiding changes in epitopes associated with adaptation during cell culture will be beneficial for production of vaccine virus.
[0009] The results presented show that Sn and CD163 work synergistically, since co-expression of both molecules results in higher virus production compared to expression of either of the two receptors alone, and this in all cell types tested. In addition, the molecular basis of this synergistic effect was elucidated, being the receptors acting at different steps during virus entry. Sn is expressed on the surface of target cells and very efficiently captures the virus and internalizes it into the cell in endosomes. CD163 on the other does not interact with the virus on the cell surface and does not internalize the virus, but co-localizes with the virus in early endosomes, where it mediates virus uncoating, followed by release of the RNA genome in the cytoplasm. Once the genome is released in the cytoplasm, genome translation, transcription and virus replication can proceed.
[0010] The finding that CD163, which is also expressed on the cell surface, does not act on the cell surface during infection, but rather interacts with the virus in endosomes inside the cell, is quite surprising. Generally, cellular receptors act during virus attachment or internalization, or direct fusion at the cell surface. This model, in which the CD163 receptor is not active on the cell surface but acts on the virus in endosomes is surprising and explains the unanticipated and cooperative action of both Sn and CD163 during virus infection, resulting in very efficient virus infection and production of high titers of virus.
[0011] Thus, these results provide new means to generate a PRRSV permissive cell that allow for efficient viral replication, with less adaptation and accordingly solves the problems recognized in the art.
[0012] This disclosure is based upon the characterization that both sialoadhesin and CD163 are not only involved in the permissivity of macrophages, the primary target cells, for a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular for PRRSV, but that these molecules also act at different steps of virus infection, thus allowing a cooperative effect during infection, resulting in enhanced virus production.
[0013] It has been found that non-permissive cells can be rendered permissive, or the permissivity of partially susceptible cells can be increased by directing the cells to express both sialoadhesin and CD163.
[0014] Provided are methods to identify the permissiveness of cells for a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular for PRRSV; the method comprising determining CD163 and sialoadhesin expression in the cells; wherein cells having a both CD163 and sialoadhesin expression, are identified as permissive cells for a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae.
[0015] Provided is a method to generate a cell(s) permissive for, or to increase the permissiveness of a cell(s) for a virus of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular for PRRSV, the method comprising treating the cells to yield an expression of both CD163 and sialoadhesin.
[0016] Further provided is a method for preparing a culture of a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, the method comprising providing a cell line identified or obtained utilzing any one of the aforementioned methods, infecting the cell line with virus and harvesting the virus from the cell culture.
[0017] Once the virus has been grown to high titres, it can be processed according to the intended use, for example in diagnosis or vaccine production, by means known in the art. For example, but not limited to, inactivating the harvested viruses with formalin, BPI, BEA or gamma-irradiation, for use in vaccines. In the alternative, the viral strain used in the infection, may be an attenuated strain for use in the production of live, attenuated vaccines.
[0018] Hence, also provided is a vaccine comprising a viral strain/serotype obtained utilzing the aforementioned method. As already mentioned hereinbefore, due to the synergetic effect of CD163 and sialoadhesin, there will be a reduction in viral adaptation and loss of altered epitopes. This taken together with the increased viral production has a tremendous effect on the antigenicity of vaccine viruses produced utilzing the methods of the disclosure. As is known to a person skilled in the art, the latter is also beneficial in isolating further viral strains from in vivo samples when diagnosing PRRSV infection in a subject.
[0019] Provided are cell lines identified or obtained utilizing any one of the aforementioned methods. Cell lines identified utilzing the methods of the disclosure include primary cell cultures and continuous cell lines obtainable thereof, but for the natural host cells, i.e., the known subpopulations of well-differentiated macrophages, in particular the alveolar macrophages that are the primary target cells of a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular of a PRRSV infection. In one embodiment, the cells consist of non-permissive PRRSV cells, such as for example PK-15, CHO, BHK-21 and Hek293t cells, expressing both Sn and CD163; within a particular embodiment, the CHO cells stably expressing sialoadhesin and CD163 deposited on May 14, 2008 at the Belgian Coordinated Collections of Microorganisms as CHO-Sn/CD163 IC5; CHO-Sn/CD163 ID9 and CHO-Sn/CD163 IF3 with the respective accession numbers LMBP 6677CB, LMBP 6678CB, and LMBP 66779 CB, respectively.
[0020] The cell lines identified or obtained utilzing any one of the aforementioned methods, can also be used in a method of diagnosing a viral infection of a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular of a PRRSV infection in a subject. Further provided is a method for diagnosing a viral infection of a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular of a PRRSV infection in a subject, the method comprising contacting a cell line identified or obtained utilzing any one of the methods of the disclosure with a sample taken from the subject and determine whether viral replication occurs.
[0021] Alternatively, the viral infection is determined by assessing the presence of virus-specific antibodies in the sample taken from the subject. In this embodiment, the cell line identified or obtained utilzing any one of the methods of the disclosure is infected with a virus of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular with PRRSV, and the reaction of the antibodies in a sample taken from the subject is done by means well known to the person skilled in the art.
[0022] In these diagnostic methods, the sample taken from the subject, is typically a biological fluid; such as for example serum, colostrums, bronchoalveolar lavage fluids, saliva, urine or feces; tissue or a tissue extract. The tissue or tissue extract to be analyzed includes those which are known, or suspected, to be permissive for the virus such as, for example PBMC (peripheral blood mononuclear cells), alveolar macrophages, lymphoid tissues such as lymph nodes, spleen, tonsils and thymus and non-lymphoid tissues such as lungs and liver.
[0023] The cell lines identified or obtained utilzing any one of the aforementioned methods can be used in a method to identify anti-viral compounds, i.e., anti-viral compounds for a virus of the family Arteriviridae or Coronaviridae or Asfarviridae as defined herein, in particular for PRRSV. Accordingly, provided is a method to identify anti-viral compounds, the method comprising contacting a cell line infected with a virus of the Arteriviridae or Coronaviridae or Asfarviridae, with the compound to be tested; and determine the capability of the test compound to modulate the viral replication in the cell line.
[0024] The capability of a compound to modulate the viral replication can be determined by utilzing amongst others, the presence of infectious viral particles in the media. The latter can be determined utilzing any one of the available protein measurement techniques and is typically determined utilzing late viral specific antibodies, in particular, utilzing (virus) specific antibodies as provided hereinafter.
[0025] In the immunoassays related thereto, the amount of viral protein produced can be quantified by any standard assay such as, for example, utilzing a luminescence assay, a chemiluminescence assay, an enzyme-multiplied immunoassay technology (EMIT) assay, a fluorescence resonance excitation transfer immunoassay (FRET) assay, an enzyme channeling immunoassay (ECIA) assay, a substrate-labeled fluorescent immunoassay (SLFIA) assay, a fluorescence polarization assay, a fluorescence protection assay, an antigen-labeled fluorescence protection assay (ALFPIA), or scintillation proximity assay (SPA).
[0026] Alternatively, the effect of the compound on viral replication is determined by assessing the virus titers in the media, by quantifying numbers of infected cells by immunocytochemistry or by utilzing a MTS cytotoxicity assay to determine the cytotoxic concentration of the viral particles in the media.
BRIEF DESCRIPTION OF THE DRAWINGS
[0027] FIG. 1: Expression of sialoadhesin and CD163 on primary alveolar macrophages. Flow cytometric analysis of macrophages stained with mAb 41D3 for porcine sialoadhesin (black curve) or mAb 2A10 for porcine CD163 (black curve). In both experiments, the isotype-matched (IgG1) antibody 13D12 (white curve) was used as control.
[0028] FIG. 2: Effect of sialoadhesin and CD163 specific antibodies on PRRSV infection of macrophages. Panel A Macrophages were treated with different concentrations of sialoadhesin and CD163 recognizing antibodies at 37° C. and inoculated with Marc-grown Lelystad virus. The relative percentage of infected macrophages was calculated, with untreated cells (RPMI) as reference. Each value represents the means±standard deviation of three experiments. Panel B Macrophages were treated with 3.3 μg/100 μl of sialoadhesin and CD163 specific antibodies and inoculated with different PRRSV strains. The relative percentage of infected macrophages was calculated, with untreated cells (RPMI) as reference. Each value represents the means±standard deviation of three experiments.
[0029] FIG. 3: PRRSV infection of non-permissive cells expressing sialoadhesin (Sn), CD163 or the combination of both. Transfected PK-15, CHO-K1 and BHK-21 cells were inoculated with either Lelystad virus or VR-2332. Twenty-four hours post-inoculation, supernatant was collected and infectious extracellular virus (black bars) was determined via titration, with 0.8 tissue culture infectious doses TCID50/ml (log10) being the detection limit. Background virus still remaining after removal of the inoculum (grey bars) was also determined. Each value represents the means±standard deviation of three experiments.
[0030] FIG. 4: Kinetics of PRRSV infection in PK-15 cells expressing sialoadhesin and CD163. PK-15 cells expressing sialoadhesin in combination with CD163 were inoculated with PRRSV at a moi of 0.1 (dashed line) or a moi of 1 (full line). At different time points after inoculation, extra- and intracellular virus was collected and titrated with 0.8 TCID50/ml (log10) being the detection limit. Each value represents the means±standard deviation of three experiments.
[0031] FIG. 5: Confocal microscopical analysis of PRRSV during infection of transfected PK-15 cells expressing PRRSV receptors sialoadhesin and/or CD163. The relative number of PK-15 cells with internalized virus particles was calculated with 1 hour post-inoculation as reference point. Data are shown for PK-15 cells expressing sialoadhesin (black bars) and the combination of sialoadhesin and CD163 (grey bars), but not for CD163-expressing PK-15 since no internalized virus particles were observed.
[0032] FIG. 6: Effect of sialoadhesin- and CD163-specific antibodies on PRRSV attachment to macrophages. Macrophages were treated with different concentrations of sialoadhesin- and CD163-specific antibodies at 4° C. and inoculated with Lelystad virus at 4° C. Unbound virus was then washed away and infection was allowed by shifting the cells to 37° C. for 10 hours. The relative percentage of infected macrophages was calculated, with untreated cells as reference. Each value represents the means±standard deviation of three experiments.
[0033] FIG. 7A: Sensitivity of CHOSn-CD163 cells to PRRSV infection. Three different densities of cells (100,000, 200,000 and 300,000 cells/ml were infected at 1, 2 or 3 days post-seeding with LV marc-grown cells. After 48 hours, cells were stained with P3/27 primary antibodies and three microscope fields (500 cells per field) were counted, and represented as the absolute amounts of infected cells for the microscopic fields.
[0034] FIG. 7B: The same as FIG. 7A, but the cells were now pretreated with neuraminidase and subsequently infected with LV macrophage grown virus.
DESCRIPTION OF THE INVENTION
[0035] The instant disclosure is based upon the observation that both sialoadhesin and CD 163 are involved in the permissivity of macrophages, the primary target cells, for a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular for PRRSV. In addition, and surprisingly, it was shown that CD163 does not act at the cell surface of susceptible cells during attachment and internalization, but rather acts during virus uncoating and genome release inside the cell in endosomes. This unexpected finding explains why CD163 acts synergistically with Sn during virus infection, since the latter interacts with the virus at the cell surface to allow virus attachment and internalization.
[0036] Provided are methods to identify the permissiveness of cells for a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular, for PRRSV, the method comprising: determining CD163 and sialoadhesin expression in the cells, wherein cells having a both CD163 and sialoadhesin expression are identified as permissive cells for a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae.
[0037] Asfarviridae is a family of icosohedral enveloped viruses whose genome consists of a single molecule of linear double-stranded DNA of about 150,000-190,000 nucleotides long. The name of the family is derived from African Swine Fever and Related Viruses. African Swine Fever Virus (ASFV) is the type species of the Asfivirus genus and is the sole member of the family. Recently, porcine CD163 polypeptide has been surmised by implication to be the cellular receptor for ASFV (Sanchez-Torres et al., 2003).
[0038] The Arteriviridae family is grouped with the Coronaviridae and Roniviridae to form the order Nidovirales. All members of the order have enveloped particles containing a single species of single-stranded RNA that encodes for a number of proteins by means of a series of nested (Latin Nido=nest) subgenomic RNAs. The family Arteriviridae contains those members with spherical virions 45-60 nm in diameter (those of the family Coronaviridae are more than 100 nm) and which infect mammals. Their genome consists of single-stranded RNA of size 12-16 kb and with a 3'-polyA tail. Two large, overlapping ORFs at the 5'-end of the genome encode the major non-structural proteins and are expressed as a fusion protein by ribosomal frameshift. Downstream are up to nine other genes, mostly or entirely encoding structural proteins, and these are expressed from a 3'-coterminal nested set of subgenomic RNAs.
[0039] Viruses of the family of Arteriviridae includes equine arteritis virus (EAV), lactate dehydrogenase-elevating virus (LDV) and simian hemorrhagic fever virus (SHFV). The Arterivirus having the greatest economic importance is PRRSV.
[0040] Thus, the methods of the disclosure are used to identify and/or modulate the permissivity of cells for a virus selected from the group consisting of ASFV, equine arteritis virus (EAV), lactate dehydrogenase-elevating virus (LDV), simian hemorrhagic fever virus (SHFV) or PRRSV, as well as variants thereof including orthologs and paralogs; in particular human orthologs. In a particular embodiment, the methods of the disclosure are used to identify and/or modulate the permissivity of cells for PRRSV.
[0041] As used herein, the terms "permissiveness of a cell(s)," "permissivity of cell(s)" and "permissive cell(s)" refers to the ability in which a particular virus, i.e., a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, can complete its replication cycle in a given cell. This in contrast to "non-permissive" cells that do not support complete replication of a virus.
[0042] "CD163" is a member of the scavenger receptor cysteine-rich (SRCR) family of transmembrane glycoproteins, and is thought to be expressed exclusively on monocytes and macrophages. One identified role of CD 163 is to inhibit oxidative tissue damage following hemolysis by consuming hemoglobin:haptoglobin complexes by endocytosis. The subsequent release of interleukin-10 and synthesis of hemeoxygenase-1 results in anti-inflammatory and cytoprotective effects. The human CD163 gene spans 35 kb on chromosome 12, and consists of 17 exons and 16 introns.
[0043] A number of isoforms of the CD163 polypeptide, including membrane bound, cytoplasmic and secreted types, are known to be generated by alternative splicing (Ritter et al., 1999). cDNA sequences that encodes a porcine CD 163 polypeptide (Genbank accession number AJ311716), a murine CD 163 polypeptide (Genbank access number AF274883), as well as multiple human variants, exemplified by Genbank access numbers AAH51281 and CAA80543, have been reported.
[0044] As used herein, the "CD163 polypeptide" is meant to be a protein encoded by a mammalian CD 163 gene, including allelic variants as well as biologically active fragments thereof containing conservative or non-conservative changes as well as artificial proteins that are substantially identical, i.e., 70%, 75%, 80%, 85%, 87%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of the aforementioned CD 163 polypeptides. In a particular embodiment, the CD 163 polypeptide is 70%, 75%, 80%, 85%, 87%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the porcine CD 163 (encoded by Genbank Accession No. AJ311716 or bankit927381 EU016226).
[0045] By analogy, the "CD163 polynucleotide" is meant to include allelic variants as well as biologically active fragments thereof containing conservative or non-conservative changes as well as any nucleic acid molecule that is substantially identical, i.e., 70%, 75%, 80%, 85%, 87%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of the aforementioned CD163 encoding polynucleotides. In a particular embodiment, the sialoadhesin polynucleotide is 70%, 75%, 80%, 85%, 87%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleic acid molecule encoding for porcine CD163 (Genbank Accession No. AJ311716 or bankit927381 EU016226).
[0046] Biologically active fragments of CD163 are meant to include fragments that retain the activity of the full length protein, such as the isoform with SwissProt accession number Q2VL90-2, the soluble form of CD163 (sCD163), or fragments containing at least 1, 2, 3, 4, 5, 6, 7, 8 or 9 of the SRCR domain(s).
[0047] "Sialoadhesin" is a lectin-like adhesion shown to bind glycoconjugate ligands in a sialic acid-dependent manner and characterized in having conserved sialic acid binding sites. It is a transmembrane glycoprotein involved in cell-cell interactions and expressed only by a subpopulation of tissue macrophages.
[0048] cDNA sequences that encodes a porcine sialoadhesin polypeptide (Genbank accession number NM--214346), a murine sialoadhesin polypeptide (Genbank access number NM--011426), as well as a human variant (Genbank access number NM--023068), have been reported.
[0049] As used herein the "sialoadhesin" polypeptide is meant to be a protein encoded by a mammalian sialoadhesin gene, including allelic variants as well as biologically active fragments thereof containing conservative or non-conservative changes as well as artificial proteins that are substantially identical, i.e., 70%, 75%, 80%, 85%, 87%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of the aforementioned sialoadhesin polypeptides. In a particular embodiment, the sialoadhesin polypeptide is 70%, 75%, 80%, 85%, 87%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the porcine sialoadhesin (encoded by Genbank Accession No. NM--214346).
[0050] By analogy, the "sialoadhesin" polynucleotide is meant to include allelic variants as well as biologically active fragments thereof containing conservative or non-conservative changes as well as any nucleic acid molecule that is substantially identical, i.e., 70%, 75%, 80%, 85%, 87%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of the aforementioned sialoadhesin encoding polynucleotides. In a particular embodiment, the sialoadhesin polynucleotide is 70%, 75%, 80%, 85%, 87%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleic acid molecule encoding for porcine sialoadhesin (Genbank Accession No. NM--214346).
[0051] Biologically active fragments of sialoadhesin are meant to include fragments that retain the activity of the full length protein, i.e., that retain the capability of binding a virus of the family of Arteriviridae includes equine arteritis virus (EAV), lactate dehydrogenase-elevating virus (LDV) and simian hemorrhagic fever virus (SHFV), in particular of binding PRRSV. Biologically active fragments include, for example, the known soluble form of sialoadhesin, fragments containing at least 1, 2, 3 or 4 of the Ig-like domain(s); in particular the N-terminal domains; more in particular consisting of the N-terminal, variable, sialic acid-binding Ig-like domain.
[0052] As used herein, the terms "polynucleotide" and "nucleic acid" are used interchangeably to refer polynucleotides of any length, either deoxyribonucleotides or ribonucleotides or analogs (e.g., inosine, 7-deazaguanosine, etc.) thereof "Oligonucleotides" refer to polynucleotides of less than 100 nucleotides in length, preferably less than 50 nucleotides in length, and most preferably about 10 to 30 nucleotides in length. Polynucleotides can have any three-dimensional structure and may perform any function, known or unknown. The following are non-limiting examples of polynucleotides: a gene or gene fragment (for example, a probe, primer, EST or SAGE tag), exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes and primers. A polynucleotide can include modified nucleotides, such as methylated nucleotides and nucleotide analogs. If present, modifications to the nucleotide structure can be imparted before or after assembly of the polymer. The sequence of nucleotides can be interrupted by non-nucleotide components. A polynucleotide can be further modified after polymerization, such as by conjugation with a labeling component. The term also refers to both double- and single-stranded molecules. Unless otherwise specified or required, any embodiment of this invention that is a polynucleotide encompasses both the double-stranded form and each of two complementary single-stranded forms known or predicted to make up the double-stranded form.
[0053] "Polypeptide" refers to any peptide or protein comprising amino acids joined to each other by peptide bonds or modified peptide bonds. "Polypeptide" refers to both short chains, commonly referred to as peptides, oligopeptides or oligomers, and to longer chains, generally referred to as proteins. Polypeptides may contain amino acids other than the 20 gene-encoded amino acids.
[0054] "Polypeptides" include amino acid sequences modified either by natural processes, such as post-translational processing, or by chemical modification techniques which are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature.
[0055] Modifications may occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains and/or the amino or carboxyl termini. It will be appreciated that the same type of modification may be present to the same or varying degrees at several sites in a given polypeptide. Also, a given polypeptide may contain many types of modifications (see, for instance, Proteins-Structure and Molecular Properties, 2nd Ed., T. E. Creighton, W.H. Freeman and Company, New York, 1993; F. Wold, "Post-translational Protein Modifications: Perspectives and Prospects," pgs. 1-12 in Post-translational Covalent Modification of Proteins, B. C. Johnson, Ed., Academic Press, New York, 1983; Seifter et al., "Analysis for protein modifications and nonprotein cofactors," Meth. Enzymol. (1990) 182:626-646; and Rattan et al., "Protein Synthesis: Post-translational Modifications and Aging," Ann. N.Y. Acad. Sci. (1992) 663:4842).
Sequence Identity
[0056] The percentage identity of nucleic acid and polypeptide sequences can be calculated utilzing commercially available algorithms which compare a reference sequence with a query sequence. The following programs (provided by the National Center for Biotechnology Information) may be used to determine homologies/identities: BLAST, gapped BLAST, BLASTN and PSI-BLAST, which may be used with default parameters.
[0057] The algorithm GAP (Genetics Computer Group, Madison, Wis.) uses the Needleman and Wunsch algorithm to align two complete sequences that maximizes the number of matches and minimizes the number of gaps. Generally, the default parameters are used, with a gap creation penalty=12 and gap extension penalty=4.
[0058] Another method for determining the best overall match between a nucleic acid sequence or a portion thereof, and a query sequence is the use of the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci., 6:237-245 (1990)). The program provides a global sequence alignment. The result of the global sequence alignment is in percent identity. Suitable parameters used in a FASTDB search of a DNA sequence to calculate percent identity are: Matrix=Unitary, k-tuple=4, Mismatch penalty=1, Joining Penalty=30, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty=0.05, and Window Size=500 or query sequence length in nucleotide bases, whichever is shorter. Suitable parameters to calculate percent identity and similarity of an amino acid alignment are: Matrix=PAM 150, k-tuple=2, Mismatch Penalty=1, Joining Penalty=20, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty=0.05, and Window Size=500 or query sequence length in nucleotide bases, whichever is shorter.
CD163 and Sialoadhesin Expression
[0059] The "expression" generally refers to the process by which polynucleotides are transcribed into mRNA and/or the process by which the mRNA is subsequently translated into peptides, polypeptides or proteins. Hence the "expression" of a gene product, in the disclosure of CD163 and sialoadhesin, can be determined either at the nucleic acid level or the protein level.
[0060] Detection can be by any appropriate method, including, e.g., detecting the quantity of mRNA transcribed from the gene or the quantity of nucleic acids derived from the mRNA transcripts. Examples of nucleic acids derived from an mRNA include a cDNA produced from the reverse transcription of the mRNA, an RNA transcribed from the cDNA, a DNA amplified from the cDNA, an RNA transcribed from the amplified cDNA, and the like. In order to detect the level of mRNA expression, the amount of the derived nucleic acid should be proportional to the amount of the mRNA transcript from which it is derived. The mRNA expression level of a gene can be detected by any method, including hybridization (e.g., nucleic acid arrays, Northern blot analysis, etc.) and/or amplification procedures according to methods widely known in the art. For example, the RNA in or from a sample can be detected directly or after amplification. Any suitable method of amplification may be used. In one embodiment, cDNA is reversed transcribed from RNA, and then optionally amplified, for example, by PCR. After amplification, the resulting DNA fragments can for example, be detected by agarose gel electrophoresis followed by visualization with ethidium bromide staining and ultraviolet illumination. A specific amplification of differentially expressed genes of interest can be verified by demonstrating that the amplified DNA fragment has the predicted size, exhibits the predicated restriction digestion pattern and/or hybridizes to the correct cloned DNA sequence.
[0061] In hybridization methods, a probe, i.e., nucleic acid molecules having at least ten nucleotides and exhibiting sequence complementarity or homology to the nucleic acid molecule to be determined, are used. It is known in the art that a "perfectly matched" probe is not needed for a specific hybridization. A probe useful for detecting mRNA is at least about 80%, 85%, 90%, 95%, 97% or 99% identical to the homologous region in the nucleic acid molecule to be determined. In one aspect, a probe is about 50 to about 75, nucleotides or, alternatively, about 50 to about 100 nucleotides in length. These probes can be designed from the sequence of full length genes. In certain embodiments, it will be advantageous to employ nucleic acid sequences as described herein in combination with an appropriate label for detecting hybridization and/or complementary sequences. A wide variety of appropriate labels, markers and/or reporters are known in the art, including fluorescent, radioactive, enzymatic or other ligands, such as avidin/biotin, which are capable of giving a detectable signal. One can employ a fluorescent label or an enzyme tag, such as urease, alkaline phosphatase or peroxidase, instead of radioactive or other environmental undesirable reagents. In the case of enzyme tags, colorimetric indicator substrates are known that can be employed to provide a signal that is visible to the human eye or spectrophotometrically, to identify specific hybridization with complementary nucleic acid-containing samples.
[0062] Detection of the level of gene expression can also include detecting the quantity of the polypeptide or protein encoded by the gene. A variety of techniques are available in the art for protein analysis. They include but are not limited to radioimmunoassay (RIA), ELISA (enzyme linked immunoradiometric assays), "sandwich" immunoassays, immunoradiometric assays, in situ immunoassays (using e.g., colloidal gold, enzyme or radioisotope labels), western blot analysis, immunoprecipitation assays, immunofluorescent assays and PAGE-SDS. One method to determine protein level involves (a) providing a biological sample containing polypeptides; and (b) measuring the amount of any immunospecific binding that occurs between an antibody reactive to the expression product of a gene of interest and a component in the sample, in which the amount of immunospecific binding indicates the level of the expressed proteins. Antibodies that specifically recognize and bind to the protein products of these genes are required for these immunoassays. These may be purchased from commercial vendors or generated and screened utilzing methods well known in the art. See, e.g., Sambrook, Fritsch and Maniatis, Molecular Cloning: A Laboratory Manual, 2nd edition (1989); Current Protocols In Molecular Biology (F. M. Ausubel et al. eds., (1987)); the series Methods In Enzymology(Academic Press, Inc.): PCR 2: A Practical Approach (M. J. MacPherson, B. D. Hames and G. R. Taylor eds. (1995)); Harlow and Lane, eds. (1988) Antibodies, A Laboratory Manual and Animal Cell Culture (R. I. Freshney, ed. (1987)).
[0063] Provided is a method to generate a cell (or cells) permissive for, or to increase the permissiveness of a cell(s) for a virus of the family Asfarviridae or Arteriviridae, in particular for PRRSV, the method comprising treating the cells to yield an expression of both CD163 and sialoadhesin.
[0064] CD 163 and sialoadhesin expression may be facilitated or increased by methods that involve the introduction of exogenous nucleic acid into the cell. Such a cell may comprise a polynucleotide or vector in a manner that permits expression of an encoded CD 163 or sialoadhesin polypeptide.
[0065] Polynucleotides that encode CD 163 or sialoadhesin may be introduced into the host cell as part of a circular plasmid, or as linear DNA comprising an isolated protein-coding region, or in a viral vector. Methods for introducing exogenous nucleic acid into the host cell well known and routinely practiced in the art include transformation, transfection, electroporation, nuclear injection, or fusion with carriers such as liposomes, micelles, ghost cells, and protoplasts. Host cell systems hereof include plant, invertebrate and vertebrate cells systems. Hosts may include, but are not limited to, the following: insect cells, porcine kidney (PK) cells, feline kidney (FK) cells, swine testicular (ST) cells, African green monkey kidney cells (MA-104, MARC-145, VERO, and COS cells), Chinese hamster ovary (CHO) cells, baby hamster kidney cells, human 293 cells, and murine 3T3 fibroblasts. Insect host cell culture systems may also be used for the expression of the polypeptides hereof. In another embodiment, the polypeptides are expressed utilzing a drosophila expression system. Alternatively the polypeptides are expressed utilzing plant-based production platforms such as for example described in R. M. Twyman et al., Molecular farming in plants: host systems and expression technology, Trends Biotechnol. 21:570-578.
[0066] The choice of a suitable expression vector for expression of the polypeptides hereof depends upon the specific host cell to be used, and is within the skill of the ordinary artisan. Examples of suitable expression vectors include pSport and pcDNA3 (Invitrogen), pCMV-Script (Stratagene), and pSVL (Pharmacia Biotech). Expression vectors for use in mammalian host cells may include transcriptional and translational control sequences derived from viral genomes. Commonly used promoter sequences and modifier sequences which may be used in the disclosure include, but are not limited to, those derived from human cytomegalovirus (CMV), Rous sarcoma virus (RSV), Adenovirus 2, Polyoma virus, and Simian virus 40 (SV40). Methods for the construction of mammalian expression vectors are disclosed, for example, in Okayama and Berg (Mol. Cell. Biol. 3:280 (1983)); Cosman et al. (Mol. Immunol. 23:935 (1986)); Cosman et al. (Nature 312:768 (1984)); EP-A-0367566; and WO 91/18982.
[0067] Because CD163 sequences are known to exist in cells from various species, the endogenous gene may be modified to permit, or increase, expression of the CD 163 polypeptide. Cells can be modified (e.g., by homologous recombination) to provide increased expression by replacing, in whole or in part, the naturally occurring CD163 promoter with all or part of a heterologous promoter, so that the cells express CD 163 polypeptide at higher levels. The heterologous promoter is inserted in such a manner that it is operatively linked to endogenous CD163 encoding sequences. (See, for example, PCT International Publication No. WO 94/12650, PCT International Publication No. WO 92/20808, and PCT International Publication No. WO 91/09955.) It is also contemplated that, in addition to heterologous promoter DNA, amplifiable marker DNA (e.g., ada, dhfr, and the multifunctional cad gene, which encodes for carbamyl phosphate synthase, aspartate transcarbamylase, and dihydroorotase) and/or intron DNA may be inserted along with the heterologous promoter DNA. If linked to the CD 163 coding sequence, amplification of the marker DNA by standard selection methods results in co-amplification of the CD163 coding sequences in the cells.
[0068] CD163 expression may also be induced by chemical treatment. Phorbol esters, especially phorbol myristyl acetate (PMA), activate one or more isozymes of the ubiquitous membrane receptor, protein kinase C (PKC) and are particularly preferred means of increasing CD163 expression. Other methods of intracellular calcium mobilization are also contemplated.
[0069] Sialoadhesin expression may also be induced by chemical treatment. It has been reported that IFN-α does increase and is even capable to induce sialoadhesin expression in the monocyte-macrophage lineage of cells. Thus, IFN-α treatment is an alternative means of increasing/inducing sialoadhesin expression in a cell.
Cell Lines
[0070] The cell lines identified or obtained utilzing the methods of the disclosure are part of the disclosure. In one embodiment, these cell lines consist of primary cell cultures of cells identified as being permissive for a virus of the family Asfarviridae or Arteriviridae, in particular for PRRSV, provided the cells are not the natural host cells, i.e., the known subpopulations of well-differentiated macrophages, in particular the alveolar macrophages that are the primary target cells of a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular of a PRRSV infection.
[0071] Cells that are cultured directly from an animal or person are known as "primary cells." With the exception of some derived from tumors, most primary cell cultures have limited lifespan. After a certain number of population doublings cells undergo the process of senescence and stop dividing, while generally retaining viability. Methods for growing suspension and adhesion cultures of primary cells are known to the person skilled in the art, such as for example described in General Techniques of Cell Culture, Maureen A. Harrison and Ian F. Rae, Cambridge University Press 2007.
[0072] As used in the methods of the disclosure, the "primary cells" are derived from; cells that support the replication of the viruses, in particular of PRRSV. In one embodiment, the cells consist of the known subpopulations of differentiated cells of the monocyte/macrophage lineage, in particular the alveolar macrophages that are the primary target cells of a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular of a PRRSV infection. In particular embodiments of the disclosure, the "primary cells" are derived from the alveolar macrophages that are the primary target cells
[0073] This in contrast to "continuous cells" also known as "an established" or "immortalized" cell line that has acquired the ability to proliferate indefinitely either through random mutation or deliberate modification, such as artificial expression of the telomerase gene. There are numerous well established cell lines representative of particular cell types.
[0074] In the context of the disclosure, the continuous cells are either derived by immortalization from the primary cell cultures mentioned herein before or obtained from a well established continuous cell line treated, in particular by transfection with a nucleic acid sequence encoding CD163 and/or sialoadhesin, to yield a stable expression of both CD163 and sialoadhesin. In an alternative embodiment, the CD163 expression may also be induced by chemical treatment in continuous cell lines stably expressing Sn. Phorbol esters, especially phorbol myristyl acetate (PMA), activate one or more isozymes of the ubiquitous membrane receptor, protein kinase C (PKC) and are particularly preferred means of increasing CD163 expression. Other methods of intracellular calcium mobilization are also contemplated. In analogy, continuous cells stably expressing CD163 may also be induced to express Sn by chemical treatment, utilzing for example IFN-α.
[0075] Examples of continuous cell lines derived from the monocyte/macrophage lineage and useful in the context of the disclosure are THP-1, MM-1, J774, SU-DHL, RAW264, 3D4, and others.
[0076] Several established methods exist for immortalizing mammalian cells in culture. Viral genes, including Epstein-Barr virus (EBV), Simian virus 40 (SV40) T antigen, adenovirus E1A and E1B, and human papillomavirus (HPV) E6 and E7 can induce immortalization by a process known as viral transformation. Although the process is reliable and relatively simple, these cells may become genetically unstable (aneuploid) and lose the properties of primary cells. For the most part, these viral genes achieve immortalization by inactivating the tumor suppressor genes that put cells into a replicative senescent state. The preferred method to immortalize cells is through expression of the telomerase reverse transcriptase protein (TERT), particularly those cells most affected by telomere length (e.g., human). This protein is inactive in most somatic cells, but when hTERT is exogenously expressed the cells are able to maintain telomere lengths sufficient to avoid replicative senescence. Analysis of several telomerase-immortalized cell lines has verified that the cells maintain a stable genotype and retain critical phenotypic markers.
[0077] The well established continuous cells used herein are typically selected from the group consisting of cells with leukocyte characteristics or non-leukocyte cells such as for example swine testicle cells, swine kidney cells (e.g., PK15 (CCL-33), SK-RST (CRL-2842)), epithelial cell cultures, skin keratinocytes (e.g., HEK001 (CRL-2404), CCD1102 (CRL-2310)), Vero cells (CCL-81), human fetal lung fibroblasts (e.g., HFL1 (CCL-153)), human embryonic lung cells (e.g., HEL299 (CCL-137)), Chinese Hamster Ovary cells (CHO) or human embryonic kidney cells (HEK).
[0078] Hence, in a further embodiment, the cell lines consist of continuous cells expressing both CD163 and sialoadhesin.
Virus Culture and Vaccines
[0079] Provided is a method for preparing a culture of a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, the method comprising providing a cell line identified or obtained utilzing any one of the aforementioned methods, infecting the cell line with virus and harvesting the virus from the cell culture.
[0080] Once the virus has been grown to high titers, it can be processed according to the intended use, for example in diagnosis or vaccine production, by means known in the art.
[0081] For example, in case of vaccine production, the harvested viruses may be inactivated for example with formalin, BPI, BEA or gamma-irradiation, for use in vaccines. In the alternative, the viral strain used in the infection, may be an attenuated strain for use in the production of live, attenuated vaccines.
[0082] Hence, the disclosure also provides a vaccine comprising a viral strain/serotype obtained utilzing the aforementioned method.
[0083] Killed (inactivated) or live vaccines can be produced. To make a live vaccine, a viral isolate, or an attenuated or mutated variant thereof, is grown in cell culture. The virus is harvested according to methods well known in the art. The virus may then be concentrated, frozen, and stored at -70° C., or freeze-dried and stored at 4° C. Prior to vaccination the virus is mixed at an appropriate dosage, (which is from about 10 to 108 tissue culture infectious doses per ml), with a pharmaceutically acceptable carrier such as a saline solution, and optionally an adjuvant.
[0084] The vaccine produced may also comprise an inactivated vaccine comprising a PRRSV strain obtained by the methods hereof. The inactivated vaccine is made by methods well known in the art. For example, once the virus is propagated to high titers, the virus antigenic mass could be obtained by methods well known in the art. For example, the virus antigenic mass may be obtained by dilution, concentration, or extraction. All of these methods have been employed to obtain appropriate viral antigenic mass to produce vaccines. The virus is then inactivated by treatment with formalin, betapropriolactone (BPL), binary ethyleneimine (BEI), or other methods known to those skilled in the art. The inactivated virus is then mixed with a pharmaceutically acceptable carrier such as a saline solution, and optionally an adjuvant. Examples of adjuvants include, but not limited to, aluminum hydroxide, oil-in-water and water-in-oil emulsions, AMPHIGEN, saponins such as QuilA, and polypeptide adjuvants including interleukins, interferons, and other cytokines.
[0085] Inactivation by formalin is performed by mixing the viral suspension with 37% formaldehyde to a final formaldehyde concentration of 0.05%. The virus-formaldehyde mixture is mixed by constant stirring for approximately 24 hours at room temperature. The inactivated virus mixture is then tested for residual live virus by assaying for growth on a suitable cell line.
[0086] Inactivation by BEI is performed by mixing the viral suspension of the disclosure with 0.1 M BEI (2-bromo-ethylamine in 0.175 N NaOH) to a final BEI concentration of 1 mM. The virus-BEI mixture is mixed by constant stirring for approximately 48 hours at room temperature, followed by the addition of 1.0 M sodium thiosulfate to a final concentration of 0.1 mM. Mixing is continued for an additional two hours. The inactivated virus mixture is tested for residual live virus by assaying for growth on a suitable cell line.
[0087] Also provided is the use of a virus, of an inactivated virus or of a vaccine of the disclosure for preparing a medicament which is employed for the prophylactic and/or therapeutic treatment of PRRSV infection in animals, in particular in swine and piglets.
[0088] The vaccine used herein advantageously is provided in a suitable formulation. Preferred are such formulations with a pharmaceutically acceptable carrier. This comprises, e.g., auxiliary substances, buffers, salts, preservatives.
Diagnosis
[0089] In the diagnostic methods, the permissive cells of the disclosure are contacted with a sample taken from an infected subject; the cells cultured to allow replication of the virus; the virus harvested from the cell culture and identified utilzing art known procedures, such as for example utilzing specific antibodies for a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular for PRRSV.
[0090] The (virus-) specific antibody is in particular a monoclonal antibody or a derivative thereof, the latter preferably selected from the group of antibody fragments, conjugates or homologues, but also complexes and absorbates known to the skilled artisan. In a particular embodiment, the (virus-) specific antibodies are selected from the group consisting of PRRSV nucleocapsid-specific antibodies such as P3/27 and SDOW17 and WBE1, 4, 5 and 6.
[0091] In the alternative, the disclosure provides the use of the viruses harvested from the above mentioned cell cultures in methods to determine a viral infection or a previous viral infection in a subject, by assessing the presence of virus-specific antibodies in the sample taken from the subject, i.e., detecting the binding of the virus-specific antibodies to the viruses harvested from the cell culture or the binding of the virus-specific antibodies to the viral protein-expressing infected cells.
[0092] A variety of techniques are available in the art to determine binding of the virus-specific antibodies to the virus. They include but are not limited to radioimmunoassay (RIA), ELISA (enzyme linked immunoradiometric assays), "sandwich" immunoassays, "competition" immunoassays, immunoradiometric assays, in situ immunoassays (using, e.g., colloidal gold, enzyme or radioisotope labels), western blot analysis, immunoprecipitation assays, immunofluorescent assays and PAGE-SDS.
[0093] In one embodiment, the presence of virus-specific antibodies will be determined utilzing a typical competition or sandwich assay. For example, in a sandwich assay the binding of the virus-specific antibodies is done utilzing a secondary labeled antibody, which is reactive for the primary virus-specific antibody and preferably has the ability to react with multiple sites on the primary antibody. In a competition assay a standard amount of a labeled virus-specific antibody will compete with the antibodies present in the sample for binding to the virus.
[0094] Known labels are of the radioactive or fluorometric type, which are detected by instrumentation, and colorimetric labels, typically enzyme labels which cause the conversion of a corresponding substrate to colored form.
[0095] Enzymes have often been used as labels in immunoassay. In conventional enzyme immunoassay (EIA), an enzyme is covalently conjugated with one component of a specifically binding antigen-antibody pair, and the resulting enzyme conjugate is reacted with a substrate to produce a signal which is detected and measured. The signal may be a color change, detected with the naked eye or by a spectrophotometric technique, or may be conversion of the substrate to a product detected by fluorescence.
[0096] Also provided is a method to determine a viral infection or a previous viral infection in a subject, the method comprising:
[0097] harvesting a virus that is a member of the family Arteriviridae or Coronaviridae or Asfarviridae, in particular for PRRSV, from a cell culture obtainable utilzing any one of the methods of the disclosure;
[0098] contacting the virus with a sample taken from the subject; and
[0099] determine the presence of virus-specific antibodies in the sample taken from the subject.
[0100] In any one of the diagnostic methods, mentioned hereinbefore, the sample is typically a biological fluid; such as for example serum, colostrums, bronchoalveolar lavage fluids, saliva, urine or feces; tissue or a tissue extract. The tissue or tissue extract to be analyzed includes those which are known, or suspected, to be permissive for the virus such as, for example PBMC (peripheral blood mononuclear cells), alveolar macrophages, lymphoid tissues such as lymph nodes, spleen, tonsils and thymus and non-lymphoid tissues such as lungs and liver.
[0101] The disclosure will be better understood by reference to the following Experimental Details, but those skilled in the art will readily appreciate that these are only illustrative. Additionally, throughout this application, various patents and publications are cited. The disclosure of these is hereby incorporated by reference into this application to describe more fully the state of the art to which this invention pertains. The following examples illustrate the invention. Other embodiments will occur to the person skilled in the art in light of these examples.
EXAMPLES
Methods
Cell Culture and Transfection
[0102] Primary alveolar macrophages were obtained from 4- to 6-week old conventional Belgian Landrace pigs from a PRRSV-negative herd as described by Wensvoort et al. (Wensvoort et al., 1991), and cultivated in RPMI 1640 supplemented with 10% fetal bovine serum (FBS), 1% nonessential amino acids and 1 mM sodium pyruvate. Marc-145 cells were cultivated in Minimum Essential Medium with Earle's salts (MEM) supplemented with 5% FBS. PK-15 cells were grown in MEM supplemented with 10% FBS. BHK-21 cells were cultivated in MEM supplemented with 10% FBS, 1% nonessential amino acids and 1 mM sodium pyruvate. CHO-K1 cells were cultivated in F-12 medium supplemented with 10% FBS and 1 mM sodium pyruvate. All cells were grown in their specific medium supplemented with 2 mM L-glutamine and a mixture of antibiotics in a humified 5% CO2 atmosphere at 37° C. PK-15, BHK-21 and CHO-K1 cells were transfected respectively with lipofectamine (Invitrogen), lipofectamine 2000 (Invitrogen) and FuGENE 6 (Roche) according to the manufacturers' instructions.
Viruses
[0103] A thirteenth passage on macrophages of the European prototype PRRSV strain, Lelystad virus (LV) (kindly provided by G. Wensvoort), was used (Wensvoort et al., 1991). The European PRRSV strain was first passaged on macrophages and subsequently cultivated on Marc-145 cells for four passages, while for the American prototype PRRSV strain, VR-2332, a fourth passage on Marc-145 cells was used which was never passaged on macrophages (Collins et al., 1992). From the Belgian isolate 94v360 a fifth passage on Marc-145 cells was used (Duan et al., 1997a).
Antibodies
[0104] CD163 was detected via mouse monoclonal anti-porcine CD163 antibody (mAb) 2A10 (Ab-Direct) (Bullido et al., 1997; Sanchez et al., 1999) or goat polyclonal anti-human CD163 antibody (pAb) (R&D Systems). For porcine sialoadhesin recognition, mAb 41D3 has been used (Duan et al., 1998b; Vanderheijden et al., 2003). Isotype-matched irrelevant mAb 13D12 directed against gD of pseudorabies virus (Nauwynck and Pensaert, 1995) and purified goat antibodies were used as negative controls. PRRSV was visualized via the nucleocapsid-recognizing mAb P3/27 (Wieczorek-Krohmer et al., 1996) or a polyclonal swine serum obtained from PRRSV infected pigs.
Constructs
[0105] CD163 variants differing in their cytoplasmic tail have been described (Nielsen et al., 2006). Since these variations do not appear to determine PRRSV receptor function (Calvert et al., 2007), only one variant, the short one, has been cloned. Therefore, total cellular RNA was isolated from porcine macrophages via the RNeasy Mini Kit (Qiagen) and subsequently converted into cDNA via oligo dT primers (Invitrogen) and SuperScript II reverse transcriptase (Invitrogen) followed by an RNase H (Gibco) treatment. The obtained single stranded cDNA served on its turn as template for PCR amplification of the CD163 sequence via the Platinum Pfx polymerase (Invitrogen) and following primers: forward primer 5'CAC CAT GGA CAA ACT CAG AAT GGT GCT ACA TGA AAA CTC T3' (SEQ ID NO:15) and reverse primer 5'TCA TTG TAC TTC AGA GTG GTC TCC TGA GGG ATT 3' (SEQ ID NO:16) (Invitrogen). The PCR fragment was then finally cloned in the pcDNA3.1D/V5-His-TOPO vector (Invitrogen).
[0106] Sialoadhesin was cloned into the same vector as was described by Vanderheijden et al. (2003). All sequences were verified via restriction digestion and sequencing.
Stable Cell Lines
[0107] To construct a cell line co-expressing Sn and CD163, CHO-K1 cells were transfected with a plasmid containing the Sn cDNA and a geneticine resistance gene. After selection for geneticine resistance, cells were transfected with a plasmid containing the CD163 cDNA and a zeocin resistance gene, which allowed selection of cells expressing both Sn and CD163. Finally, 16 clones that co-expressed Sn and CD163 (CHOSn-CD163) were isolated. Ten clones in which 100% of the cells stably expressed Sn and CD163 were isolated, while the other six clones lost either Sn or CD 163 receptor expression. After a first screening for susceptibility to PRRSV infection, three CHOSn-CD163 clones, i.e., IC5, ID9 & IF3 were selected for further research and deposited at the Belgian Coordinated Collections of Microorganisms as CHO-Sn/CD163 IC5; CHO-Sn/CD163 ID9 and CHO-Sn/CD163 IF3 with the respective accession numbers LMBP 6677CB; LMBP 6678CB; and LMBP 66779 CB respectively.
Viral Inactivation
[0108] Inactivation with ultraviolet (UV) radiation was performed with a UV cross-linker (UVP, Inc). Purified virus (107 TCID50/ml) was radiated with UV light of different doses (0-100-1000-2000-3000 or 4000 mJ/cm2) (Darnell et al., 2004). Inactivation with binary ethyleneimine (BEI) was done by incubating purified virus (107 TCID50/ml) with 1 mM BEI (Aldrich) for several (0-6-12-24-48 or 72 hours) at 37° C. The reaction was stopped with 0.1 mM sodium thiosulfate (Sigma) (Mondal et al. 2005).
Flow Cytometry
[0109] Twenty-four hours after seeding, macrophages were lifted from the cell culture plate by incubation with ice-cold PBS for 30 minutes at 4° C. immediately prior to immunostaining and flow-cytometric analysis. Cells were first fixed with 3% paraformaldehyde followed by washing and incubation with primary mAb 41D3, 2A10 or isotype-matched control antibodies at 4° C. Afterwards, cells were washed three times and subsequently incubated with FITC-labeled goat-anti-mouse Ab (Molecular Probes). Finally, cells were washed three times, resuspended in phosphate buffered saline (PBS) and analyzed with a Becton-Dickinson (San Jose, Calif.) FACScalibur. Twenty thousand cells were analyzed for each sample and three parameters were stored for further analysis: forward light scatter, sideward light scatter and green fluorescence.
Virus Titration
[0110] To determine the titer of extracellular virus, supernatant was collected and centrifuged to remove cell debris. To determine the titer of intracellular virus, cells were washed, collected and lysed by three cycles of freeze-thawing. For titration on Marc-145 cells, cells were planted three days before inoculation. Then, they were inoculated with a ten-fold dilution series of the samples and incubated for seven days at 37° C. followed by evaluation of the cytopathic effect (CPE). For titration on macrophages, cells were planted one day before inoculation followed by inoculation with a ten-fold dilution series of the samples, incubation for three days at 37° C. and finally evaluation. CPE was studied and furthermore infected cells were visualized via an immunoperoxidase monolayer assay (IPMA) (Wensvoort et al., 1991).
Immunofluorescence Staining and Microscopy
[0111] Transfected and/or infected cells were fixed with ice-cold methanol, unless pictures or colocalization studies were demanded. In those cases 3% paraformaldehyde was used and if needed, those cells were permeabilized with 0.1% Triton X-100. Fixation and permeabilization reagentia were removed via three times washing with PBS. Fixed cells were incubated with primary antibodies for at least 1 hour at 37° C., washed three times with PBS and further incubated with secondary antibodies for at least 1 hour at 37° C. Finally, cells were washed three times, embedded in glycerine-DABCO, mounted and analyzed via fluorescence microscopy.
Colocalization
[0112] To quantitate colocalization between sialoadhesin and CD163 on the surface of macrophages, confocal images were taken and analyzed via the program CoLocalizer Pro. Prior to merging the two images, weak fluorescent background was substracted. Based on the overlay, different colocalization parameters were calculated according to the manual.
Treatment of Macrophages with Sialoadhesin and CD163 Specific Antibodies
[0113] Macrophages were seeded in 96-well plates 24 hours before the experiment was performed. A three-fold dilution series was prepared for different antibodies (2A10, CD163-pAb, 41D3, 13D12, purified goat antibodies) and the HbHp complex (Hb AoH0267, Hp type2-2 H9762 from Sigma-Aldrich), which was made by 15 minutes incubation of both components at room temperature. Macrophages were then incubated for one hour at 37° C. with the concentration gradient of antibodies/proteins followed by inoculation with PRRSV in the presence of antibodies/proteins for one hour at 37° C. After the treatment, cells were washed, further incubated for 9 hours at 37° C. and fixed with methanol. Infected cells were visualized via an immunoperoxidase staining with mAb P3/27 or the polyclonal swine serum as primary antibodies and respectively goat-anti-mouse HRP or rabbit-anti-swine HRP (Dako) as secondary antibodies. In control reactions, no difference in the percentage of infected cells was observed for the two different PRRSV-recognizing antibodies. After counting the percentage of infected cells, relative percentages of infection were calculated with cells without any antibody/protein treatment as reference value represented by the RPMI data.
Infection Experiments on Non-Target Cells and on Macrophages
[0114] For different infection experiments, a similar protocol was used as will be described here. Twenty-four hours post-transfection of PRRSV non-target cells or 24 hours post-seeding of the macrophages, cells were washed once with RPMI followed by inoculation with PRRSV viral supernatant which was cleared from cell debris via centrifugation. Inoculated cells were incubated for 1 hour at 37° C. in the presence of the virus. After virus removal, macrophages were directly covered with medium unlike non-target cells, which were washed five times with RPMI before incubation in medium. The final wash solution was collected and titrated to determine the amount of background virus still present after removal of the inoculum. At different time points after inoculation, cells were fixed with ice-cold methanol or paraformaldehyde and intra- and extracellular virus was collected as described in virus titration.
Infection Experiments on CHOSn-CD163 Cells
[0115] For different infection experiments, a similar protocol was used as will be described here. Three different CHOSn-CD163 cell clones (IC5, ID9 and IF3) seeded at different densities (100,000, 200,000 or 300,000 cells/ml) are infected at different days post-seeding (1, 2 or 3 days post-seeding) with LV marc grown (moi 1) or with LV macrophage grown (moi 10). After 48 hours post-inoculation the cells are fixed with methanol and stained with primary antibody P3/27 and secondary antibody goat-anti-mouse HRP. Afterwards AEC substrate is added. With a microscope infected cells of three fields with a 40× lens (500 cells per field) are counted. To enhance virus interaction with the sialoadhesin receptor that depends on interaction of virus-linked sialic acids with the N-terminal, sialic acid binding domain of Sn, and subsequent infection, the cells were treated with neuramimidae (50 mU/ml of Vibirio cholerae neuraminidase) to remove cis-acting cellular sialic acids prior to infection with the PRRSV macrophage grown viruses.
Results
1. Expression of PRRSV Receptors Sialoadhesin and CD163 on Macrophages, the In Vivo Target Cells of PRRSV
[0116] Both PRRSV receptors sialoadhesin and CD163 are described to be restricted to monocytes-macrophages (Duan et al., 1998b; Sanchez et al., 1999). To further investigate their role in PRRSV infection, we first wanted to confirm their presence and analyze their expression pattern in a population of macrophages. Therefore, macrophages were cultivated for 24 hours, lifted from the cell culture plate, immunostained and analyzed via flow cytometry or they were cultivated for 24 hours, fixed, immunostained and analyzed via confocal microscopy. Similar to previous reports, the flow cytometry data (FIG. 1A) shows that nearly 100% of the macrophages are positive for both PRRSV receptors. Thus, macrophages represent a homogenous population with respect to the expression of sialoadhesin and CD163. Immunofluorescence staining followed by confocal analysis (FIG. 1B) reveals an almost exclusive expression of sialoadhesin on the cell membrane. This is in contrast to CD163, which is clearly present on the cell membrane and also intracellularly.
2. Effect of Sialoadhesin and CD163 Specific Antibodies on PRRSV Infection of Macrophages
[0117] To examine the role of CD163 in PRRSV infection of primary macrophages, the effect of a CD163-specific mAb and a pAb on PRRSV infection of macrophages was evaluated. Therefore, macrophages were pre-incubated with different concentrations of antibodies followed by inoculation of the macrophages with PRRSV in the presence of the antibodies for 1 hour, washing, further incubation for 9 hours at 37° C., methanol-fixation and finally staining with PRRSV-specific antibodies. Relative percentages of infected macrophages are represented in FIG. 2A showing that mAb 41D3, directed against porcine sialoadhesin, strongly reduces PRRSV infection in a dose dependent manner, as has been described before (Duan et al., 1998a). Interestingly, also the CD163-specific polyclonal antibody clearly reduces PRRSV infection up to 75% in a dose dependent fashion, suggesting a role for CD163 in PRRSV infection of macrophages. However, the CD163-specific monoclonal antibody had no effect on PRRSV infection (data not shown). Also the best-characterized ligand of CD163, the hemoglobin-haptoglobin complex, did not influence PRRSV infection (data not shown). Ultimately, when 41D3 and the CD163-specific polyclonal antibody were combined, PRRSV infection was completely blocked. None of the negative controls, being irrelevant isotype-matched control antibodies, had an effect on PRRSV infection. These results clearly demonstrate that CD163 and sialoadhesin are involved in PRRSV infection of macrophages.
[0118] PRRSV displays remarkable genetic, antigenic, and clinical variability resulting in distinct groups of isolates within the same viral family (Goldberg et al., 2003), urging the need to investigate whether sialoadhesin and CD163 are involved in infection of different PRRSV strains (FIG. 2, Panels B and C). Therefore, different isolates were tested for their infectivity on macrophages in the presence of PRRSV-receptor-recognizing antibodies as described above. The European prototype PRRSV strain Lelystad virus (LV), the American prototype strain VR-2332 and the Belgian isolate 94v360 all show a clear reduction in the presence of 41D3 and the CD163-specific polyclonal antibody and an even greater reduction when both antibodies are combined. All three isolates used were adapted to grow on Marc-145 cells. Interestingly, the LV strain grown on macrophages without any adaptation to a cell-line shows the same trend, suggesting that the genetic diversity or the producer cells of PRRSV do not influence the need of different PRRSV strains for sialoadhesin and CD163 to infect macrophages.
3. PRRSV Non-Target Cells Expressing Both Sialoadhesin and CD163 Support Productive PRRSV Infection and are More Efficient Compared to CD163 Alone
[0119] Since the experiment with antibodies showed that both sialoadhesin and CD163 are involved in PRRSV infection of macrophages, we investigated their potential role in productive PRRSV infection in different non-susceptible cell-lines, either expressed separately or combined. PK-15, CHO-K1 and BHK-21 cells were transfected with sialoadhesin, CD163 or a combination of both and 24 hours post-transfection, cells were inoculated with the European prototype Lelystad virus or the American prototype VR-2332 virus. At 24 hours post-inoculation, supernatant was collected and cells were fixed. The supernatant was titrated on Marc-145 cells to determine the amount of infectious virus produced in the transiently transfected cells (FIG. 3). The fixed cells were analyzed via immunofluorescence microscopy for the presence of PRRSV.
[0120] In addition to PK-15, CHO-K1 and BHK-21 cells, HEK293t cells were tested for their susceptibility to PRRSV infection. Similar to the three other cell lines, HEK293t cells expressing sialoadhesin did not support productive PRRSV infection. HEK293t cells expressing CD163 alone supported productive PRRSV infection and 10 to 100 times more infected cells were observed in cells expressing both sialoadhesin and CD163 (data not shown).
[0121] Bottom line, cells only expressing sialoadhesin never showed infection, as was already noted by Vanderhijden et al. (2003). Infected cells were only observed when CD163 was present, alone or in combination with sialoadhesin (data not shown), but where CD163 alone is able to sustain PRRSV infection, 10 to 100 times more infected cells were observed in cells expressing both sialoadhesin and CD 163.
[0122] In agreement with the results obtained via immunofluorescence microscopy, no extracellular virus was detected for cells only expressing sialoadhesin. Except for PK-15 cells, were some extracellular virus is present, however without showing infected cells. When only CD163 is expressed, all three cell-lines produce new infectious virus, but the virus titers are rather low probably because of low infection efficiency. When both sialoadhesin and CD163 are present, all three cell-lines produce new infectious virus with virus titers remarkably higher compared to cells with only CD163, especially for PK-15 and CHO-K1 cells. Comparison between the European and the American prototype PRRSV strain shows higher virus titers for the VR-2332 strain in PK-15 and CHO-K1 cells but not in BHK-21 cells. Furthermore, for one repetition of the experiment the titration was performed not only on Marc-145 cells but also on macrophages revealing the same virus titers. Thus, PRRSV non-target cells expressing CD163 or CD163 combined with sialoadhesin produce new virus that is infectious on both Marc-145 cells and on macrophages. Further support, to the fact that non-permissive PRRSV cells can be rendered permissive with high virus titers when expression both CD163 and Sialoadhesin, was given in assessing the infectivity of the European prototype PRRSV strain Lelystad, the American prototype PRRSV strain VR-2332, and the Belgian PRRSV isolate 94V360 on the PK-15, CHO-K1, BHK-21 and HEK293t cells. For all four cell lines, similar results were observed as described for the two prototype strains. 94V360 was not able to infect cells only expressing sialoadhesin. Cells expressing CD163 were able to sustain PRRSV infection, however 10 to 100 times more infected cells were observed in cells expressing both sialoadhesin and CD163.
4. Kinetics of PRRSV Infection in PK-15 Cells Expressing Both Sialoadhesin and CD163
[0123] Because the combination of sialoadhesin and CD163 efficiently supports PRRSV infection in non-permissive cells, we wanted to study the kinetics of PRRSV infection in those non-target cells. Therefore, swine kidney PK-15 cells were transfected with a combination of sialoadhesin and CD 163 and 24 hours post-transfection, cells were inoculated with Lelystad virus at a moi of 0.1 or 1. At different time points after infection, intra- and extracellular virus was collected to be titrated on Marc-145 cells, as shown in FIG. 4. Starting from 12 hpi, an increase in the extracellular virus titer can be seen, which reaches its maximum around 48 hpi. Afterwards the titer drops which can possibly be explained by the limited number of sialoadhesin and CD163 expressing cells and/or the virus that becomes inactivated by the temperature. The amount of internalized virus particles is clearly dependent upon the titer in the inoculum. In the first six hours, the amount of internalized virus stays the same or shows a little drop. Afterwards it increases to reach a maximum around 24 hpi, which on its turn, is followed by a decrease of the intracellular virus.
5. Specific Function for Sialoadhesin and CD163 During PRRSV Infection of PK-15 Cells
[0124] Since PRRSV infection of macrophages and non-target cells is clearly dependent upon sialoadhesin and CD163, their specific molecular contributions to PRRSV infection need to be investigated. Results previously obtained in the lab indicate that sialoadhesin is important for the internalization of the virus (Vanderheijden et al., 2003). We want to confirm these results and study the role of CD163 during PRRSV infection. Therefore PK-15 cells were transfected with sialoadhesin, CD163 or a combination of both, which was followed by inoculation with Lelystad virus. At different time points after infection cells were fixed and PRRSV was visualized via immunofluorescence staining with the mAb P3/27 recognizing the PRRSV nucleocapsid protein represented in FIG. 5. Sialoadhesin expressing cells clearly internalize PRRSV virus particles, however virus disassembly does not occur at any time point and the cells do not become productively infected. Only at 24 hpi there is a small decrease in the number of cells with internalized virus particles. In CD163 expressing cells, internalized virus particles could not be observed. Surprisingly, those cells become infected and produce new infectious virus particles. PK-15 cells expressing both sialoadhesin and CD163 internalize virus particles similar to sialoadhesin expressing PK-15 cells. However, due to the presence of CD163 a clear reduction in the number of cells showing internalized virus particles is observed at 6 hpi resulting in infection at 12 hpi and even more at 24 hpi. Thus, infection of cells expressing both sialoadhesin and CD163 is much more efficient than cells only expressing CD163, as has been shown before (FIG. 3). Those results confirm the role of sialoadhesin as internalization receptor and unexpectedly show a role for CD 163 in fusion.
6. Treatment of Macrophages with Sialoadhesin- and CD163-Specific Antibodies at 4° C.
[0125] In addition to sialoadhesin, CD163 is shown to be involved during PRRSV entry in macrophages. Sialoadhesin is known as PRRSV attachment and internalization receptor. Our data suggest a role for CD163 during PRRSV uncoating, however, further research is needed to unravel its exact functioning. Therefore, we wanted to investigate whether CD 163 is involved during PRRSV attachment.
[0126] Macrophages were seeded in 96-well plates 24 hours before the experiment was performed. A three-fold dilution series was prepared for different antibodies (pAb CD163, 41D3, 13D12, purified control goat antibodies). For treatment at 4° C., macrophages were preincubated for 30 minutes at 4° C. prior to incubation for 1 hour at 4° C. with the ligands followed by inoculation with PRRSV in the presence of a new dilution series of ligands for 1 hour at 4° C. After the inoculation, cells were washed, incubated for 10 hours at 37° C. and fixed with methanol. Infected cells were visualized via an immunoperoxidase staining with mAb P3/27 or the polyclonal swine serum as primary antibodies and respectively HRP-labeled goat-anti-mouse or rabbit-anti-swine (Dako) as secondary antibodies. No difference in the percentage of infected cells was observed for the two different PRRSV-recognizing antibodies. Cells without ligand treatment are represented as control. For these untreated cells the average percentage of infected cells was calculated from six replicates. This average percentage was used as reference value in the calculation of the relative percentages of infection.
[0127] Monoclonal antibody 41D3 reduced PRRSV infection, contrasting with the pAb directed against CD163, which did not reduce PRRSV infection when administered at 4° C. (FIG. 6). These data confirm the role of sialoadhesin as PRRSV attachment receptor and suggest that CD163 is not involved in PRRSV attachment to macrophages.
7. PRRSV Entry in Macrophages: Colocalization Between CD163 and PRRSV
[0128] CD163 is shown to be involved in PRRSV entry of macrophages, however not during attachment. Furthermore, CD163 enables PRRSV to disassemble and productively infect non-target cells, suggesting that CD163 acts during PRRSV entry. To test this hypothesis, we investigated the entry of PRRSV in macrophages via confocal analysis of immunofluorescence experiments showing PRRSV and CD163 at different time points after inoculation.
[0129] Twenty-four hours after seeding, macrophages were washed and incubated with PRRSV for 5, 10 or maximum 15 minutes at 37° C. Fifteen minutes post-inoculation, the virus was replaced by medium and cells were further incubated at 37° C. At different time point post-inoculation, cells were washed, fixed with paraformaldehyde, permeablized with TX-100 and stained. Cells were first incubated with a mAb directed against GP5 (isotype IgG2a), followed by incubation with the secondary antibody goat-anti-mouse Texas Red. Cells were then again incubated with the GP5 recognizing antibody. Finally, CD163 was visualized via mAb 2A10, which was directly labeled with Alexa 488 via the mouse IgG1 specific zenon labelings kit (Invitrogen). Stainings were analyzed via confocal microscopy.
[0130] PRRSV attaches to macrophages from 5 minutes post-inoculation and first internalized virions were observed starting from 10 minutes post-inoculation (data not shown). PRRSV bound to the surface of the macrophages did not colocalize with CD163. However, internalized virus particles ended up in CD163 positive endosomes. Starting from 45 minutes after inoculation, endosomes disappeared and PRRSV and CD163 started to separate.
[0131] The surprising observation that PRRSV and CD163 colocalize in endosomes, this in contrast to Sn which colocalizes with PRRSV on the cell surface, further shows that CD163 is not involved in PRRSV attachment to macrophages, but rather has a role in virus fusion and uncoating inside the cell in endosomes.
8. Binding, Internalization, Fusion and Infection of PRRSV in CHO Cells Stably Expressing Sn and CD163.
[0132] From Day 2 onward, there is little difference in the degree of infectivity of the stable CHOSn-CD163 cells irrespective of the fact whether the PRRSV was grown on Marc-145 cells (FIG. 7A) or macrophages (FIG. 7B). Pretreatment of the cells with neuraminidase enhanced the infection of the cells with macrophage grown PRRS virus. True permissivity of the stable CHOSn-CD163 cells was confirmed by immunostaining in the cells.
[0133] Three different CHOSn-CD163 cell clones (IC5, ID9 and IF3) were seeded at 200 000 cells/ml in a 24-well plate with insert. After two days, the cells were inoculated with LV grown on marc cells or with inactivated LV grown on marc cells. The cells were fixed with methanol after 1 hour at 4° C. (binding, at 4° C. virus is not able to internalize), 1 hour at 37° C. (internalization), 5 hours at 37° C. (fusion, this means that virus particles are dismantled, as a consequence virus staining disappears), 12 hours at 37° C. (infection) and 24 hours at 37° C. (infection). The virus was stained with a primary antibody P3/27 and a secondary antibody goat-anti-mouse FITC. The virus particles were counted with a confocal microscope.
TABLE-US-00001 TABLE 1 Binding, internalization, fusion and infection of LV (control) and inactivated LV in three different CHOSn-CD163 cell lines (IC5, ID9 and IF3) Control BEI UV IC5 ID9 IF3 IC5 ID9 IF3 IC5 ID9 IF3 Binding 14 12 7 18 9 10 5 6 5 (particles) Internalization 41 31 31 38 34 25 35 23 24 (particles) Fusion 3 2 2 1 1 2 0 0 0 (particles) Infection 0 0 0 0 0 0 0 0 0 12 hpi (%) Infection 5 2 2 0 0 0 0 0 0 24 hpi (%)
[0134] As shown in the aforementioned table, LV marc grown virus can perform a complete virus cycle in the three CHOSn-CD163 cell lines. First, the virus particles bind to the cells, then virus particles enter the cells, after which the particles are dismantled to release the genome. Finally infection occurs. Clearly, this shows that the infection route of PRRSV in these cells mimics the infection observed in the in vivo target cells, the macrophages. LV inactivated with BEI and UV show binding, internalization and fusion identical to the non-inactivated LV, but there is no infection, so these methods are good candidates for vaccine development. These results clearly demonstrate the use of the stable CHOSn-CD163 cell lines, in studying and optimizing viral inactivation processes as part of vaccine production, and accordingly provide an interesting alternative for the less accessible natural host cells.
9. Intracellular and Extracellular Virus Production on Cells Infected Two Days Post-Seeding.
[0135] Further evidence for the permissivity of the stable CHOSn-CD163 cells was shown when analyzing the intracellular and extracellular virus production in the cells by titrating the produced virus not only on the CHOSn-CD163 cells, but also on the natural host (alveolar macrophages) and the Marc-145 cells known to sustain PRRSV infection. CHOSn-CD163 clone IC5 was infected two days post-seeding with LV grown on marc cells, VR grown on marc cells, 94V360 grown on macrophages, similar to the infection procedures described hereinbefore. Pretreatment of the cells with neuraminidase was also included for infection with PRRSV 94V360 grown on macrophages. Titration of extracellular (extra) and intracellular (intra) was done at three days (Table 2) and five days (Table 3) post-inoculation, respectively, and expressed as log10 units of the TCID50/ml.
TABLE-US-00002 TABLE 2 Titration on different cells types (Marc-145, macrophages and CHOSn-CD163 clone IC5) of intra - and extra-cellular virus production 3 days post-inoculation. 94V360 LV VR 94V360 macrophage + marc marc macrophage neuraminidase Extra on marc cells 4.3 4.8 / 3.8 Extra on macrophages 4.3 5.3 4.8 5.3 Extra on CHOSn-CD163 3.3 4.8 2.8 4.3 Intra on marc cells 4.8 5.8 2.1 3.3 Intra on macrophages 4.3 5.3 3.8 4.8 Intra on CHOSn-CD163 3.8 5.3 / 3.8
TABLE-US-00003 TABLE 3 Titration on different cells types (Marc-145, macrophages and CHOSn-CD163 clone IC5) of intra- and extra-cellular virus production five days post-inoculation. 94V360 LV VR 94V360 macrophage + marc marc macrophage neuraminidase Extra on marc cells 4.3 5.3 / 3.8 Extra on macrophages 5.55 5.3 4.8 5.3 Extra on CHOSn-CD163 2.1 4.8 3.3 4.8 Intra on marc cells 3.1 4.0 2.1 2.8 Intra on macrophages 3.8 4.3 4.3 3.6 Intra on CHOSn-CD163 2.8 4.3 3.3 3.3
[0136] Together, these results clearly show that virus is produced on CHOSn-CD163 cells and that this virus can infect not only CHOSn-CD163 cells, but also Marc-145 cells and primary macrophages, showing that no virus adaptation had occurred during infection of CHOSn-CD163 cells. In particular, it is interesting that for all the viruses produced in CHOSn-CD163 cells, highest levels were always detected by titration on macrophages, again showing that no adaptation during in vitro culture had occurred that would modify virus epitopes involved in infection of macrophages and induction of neutralizing antibodies.
[0137] In conclusion these results show that the CHOSn-CD163 cells can be used for virus production for use in vaccines or diagnosis.
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[0164] Wissink E. H., H. A. van Wijk, J. M. Pol, G. J. Godeke, P. A. van Rijn, P. J. Rottier, and J. Meulenberg (2003). Identification of porcine alveolar macrophage glycoproteins involved in infection of porcine respiratory and reproductive syndrome virus. Archives of Virology 148:177-187.
Sequence CWU
1
SEQUENCE LISTING
<160> NUMBER OF SEQ ID NOS: 16
<210> SEQ ID NO 1
<211> LENGTH: 3400
<212> TYPE: DNA
<213> ORGANISM: Sus scrofa
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/AJ311716
<309> DATABASE ENTRY DATE: 2005-04-15
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(3400)
<400> SEQUENCE: 1
atggtgctac ttgaagactc tggatctgca gactttagaa gatgttctgc ccatttaagt 60
tccttcactt ttgctgtagt cgctgttctc agtgcctgct tggtcactag ttctcttgga 120
ggaaaagaca aggagctgag gctaacgggt ggtgaaaaca agtgctctgg aagagtggag 180
gtgaaagtgc aggaggagtg gggaactgtg tgtaataatg gctgggacat ggatgtggtc 240
tctgttgttt gtaggcagct gggatgtcca actgctatca aagccactgg atgggctaat 300
tttagtgcag gttctggacg catttggatg gatcatgttt cttgtcgagg gaatgagtca 360
gctctctggg actgcaaaca tgatggatgg ggaaagcata actgtactca ccaacaggat 420
gctggagtaa cctgctcaga tggatctgat ttagagatga ggctggtgaa tggaggaaac 480
cggtgcttag gaagaataga agtcaaattt caagagcggt ggggaacagt gtgtgatgat 540
aacttcaaca taaatcatgc ttctgtggtt tgtaaacaac ttgaatgtgg aagtgctgtc 600
agtttctctg gttcagctaa ttttggagaa ggttctggac caatctggtt tgatgatctt 660
gtatgcaatg gaaatgagtc agctctctgg aactgcaaac atgaaggatg gggaaagcac 720
aattgcgatc atgctgagga tgctggagtg atttgcttaa atggagcaga cctgaaactg 780
agagtggtag atggactcac tgaatgttca ggaagattgg aagtgaaatt ccaaggagaa 840
tggggaacaa tctgtgatga tggctgggat agtgatgatg ccgctgtggc atgtaagcaa 900
ctgggatgtc caactgctgt cactgccatt ggtcgagtta acgccagtga gggaactgga 960
cacatttggc ttgacagtgt ttcttgccat ggacacgagt ctgctctctg gcagtgtaga 1020
caccatgaat ggggaaagca ttattgcaat cataatgaag atgctggtgt gacatgttct 1080
gatggatcag atctggaact gagacttaaa ggtggaggca gccactgtgc tgggacagtg 1140
gaggtggaaa ttcagaaact ggtaggaaaa gtgtgtgata gaagctgggg actgaaagaa 1200
gctgatgtgg tttgcaggca gctgggatgt ggatctgcac tcaaaacatc atatcaagtt 1260
tattccaaaa ccaaggcaac aaacacatgg ctgtttgtaa gcagctgtaa tggaaatgaa 1320
acttctcttt gggactgcaa gaattggcag tggggtggac ttagttgtga tcactatgac 1380
gaagccaaaa ttacctgctc agcccacagg aaacccaggc tggttggagg ggacattccc 1440
tgctctggtc gtgttgaagt acaacatgga gacacgtggg gcaccgtctg tgattctgac 1500
ttctctctgg aggcggccag cgtgctgtgc agggaactac agtgcggcac tgtggtttcc 1560
ctcctggggg gagctcactt tggagaagga agtggacaga tctgggctga agaattccag 1620
tgtgaggggc acgagtccca cctttcactc tgcccagtag caccccgccc tgacgggaca 1680
tgtagccaca gcagggacgt cggcgtagtc tgctcaagat acacacaaat ccgcttggtg 1740
aatggcaaga ccccatgtga aggaagagtg gagctcaaca ttcttgggtc ctgggggtcc 1800
ctctgcaact ctcactggga catggaagat gcccatgttt tatgccagca gcttaaatgt 1860
ggagttgccc tttctatccc gggaggagca ccttttggga aaggaagtga gcaggtctgg 1920
aggcacatgt ttcactgcac tgggactgag aagcacatgg gagattgttc cgtcactgct 1980
ctgggcgcat cactctgttc ttcagggcaa gtggcctctg taatctgctc agggaaccag 2040
agtcagacac tatccccgtg caattcatca tcctcggacc catcaagctc tattatttca 2100
gaagaaagtg gtgttgcctg catagggagt ggtcaacttc gcctggtcga tggaggtggt 2160
cgttgtgctg ggagagtaga ggtctatcct ggggcatcct ggggcaccat ctgtgatgac 2220
agctgggacc tgaatgatgc ccatgtggtg tgcaaacagc tgagctgtgg atgggccatt 2280
aatgccactg gttctgctca ttttggggaa ggaacagggc ccatttggct ggatgagata 2340
aactgtaatg gaaaagaatc tcatatttgg caatgccact cacatggttg ggggcggcac 2400
aattgcaggc ataaggagga tgcaggagtc atctgctcag agttcatgtc tctgagactg 2460
atcagtgaaa acagcagaga gacctgtgca gggcgcctgg aagtttttta caacggagct 2520
tggggcagcg ttggcaggaa tagcatgtct ccagccacag tgggggtggt atgcaggcag 2580
ctgggctgtg cagacagagg ggacatcagc cctgcatctt cagacaagac agtgtccagg 2640
cacatgtggg tggacaatgt tcagtgtcct aaaggacctg acacactatg gcagtgcccc 2700
tcatctccat ggaagaagag actggccagc ccctcagagg agacatggat cacatgtgcc 2760
aacaaaataa gacttcaaga aggaaacact aattgttctg gacgtgtgga gatctggtac 2820
ggaggttcct ggggcactgt gtgtgacgac tcctgggacc ttgaagatgc tcaggtggtg 2880
tgccgacagc tgggctgtgg ctcagctttg gaggcaggaa aagagcccgc atttggccag 2940
gggactgggc ccatatggct caatgaagtg aagtgcaagg ggaatgaacc ctccttgtgg 3000
gattgtcctg ccagatcctg gggccacagt gactgtggac acaaggagga tgctgctgtg 3060
acgtgctcag aaattgcaaa gagccgagaa tccctacatg ccacaggtcg ctcatctttt 3120
gttgcacttg caatctttgg ggtcattctg ttggcctgtc tcatcgcatt cctcatttgg 3180
actcagaagc gaagacagag gcagcggctc tcagttttct caggaggaga gaattctgtc 3240
catcaaattc aataccggga gatgaattct tgcctgaaag cagatgaaac ggatatgcta 3300
aatccctcag gagaccactc tgaagtacaa tgaaaaggaa aatgggaatt ataacctggt 3360
gagttcagcc tttaagatac cttgatgaag acctggacta 3400
<210> SEQ ID NO 2
<211> LENGTH: 1115
<212> TYPE: PRT
<213> ORGANISM: Sus scrofa
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/CAC84397
<309> DATABASE ENTRY DATE: 2005-04-15
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1115)
<400> SEQUENCE: 2
Met Asp Lys Leu Arg Met Val Leu His Glu Asn Ser Gly Ser Ala Asp
1 5 10 15
Phe Arg Arg Cys Ser Ala His Leu Ser Ser Phe Thr Phe Ala Val Val
20 25 30
Ala Val Leu Ser Ala Cys Leu Val Thr Ser Ser Leu Gly Gly Lys Asp
35 40 45
Lys Glu Leu Arg Leu Thr Gly Gly Glu Asn Lys Cys Ser Gly Arg Val
50 55 60
Glu Val Lys Val Gln Glu Glu Trp Gly Thr Val Cys Asn Asn Gly Trp
65 70 75 80
Asp Met Asp Val Val Ser Val Val Cys Arg Gln Leu Gly Cys Pro Thr
85 90 95
Ala Ile Lys Ala Thr Gly Trp Ala Asn Phe Ser Ala Gly Ser Gly Arg
100 105 110
Ile Trp Met Asp His Val Ser Cys Arg Gly Asn Glu Ser Ala Leu Trp
115 120 125
Asp Cys Lys His Asp Gly Trp Gly Lys His Asn Cys Thr His Gln Gln
130 135 140
Asp Ala Gly Val Thr Cys Ser Asp Gly Ser Asp Leu Glu Met Gly Leu
145 150 155 160
Val Asn Gly Gly Asn Arg Cys Leu Gly Arg Ile Glu Val Lys Phe Gln
165 170 175
Gly Arg Trp Gly Thr Val Cys Asp Asp Asn Phe Asn Ile Asn His Ala
180 185 190
Ser Val Val Cys Lys Gln Leu Glu Cys Gly Ser Ala Val Ser Phe Ser
195 200 205
Gly Ser Ala Asn Phe Gly Glu Gly Ser Gly Pro Ile Trp Phe Asp Asp
210 215 220
Leu Val Cys Asn Gly Asn Glu Ser Ala Leu Trp Asn Cys Lys His Glu
225 230 235 240
Gly Trp Gly Lys His Asn Cys Asp His Ala Glu Asp Ala Gly Val Ile
245 250 255
Cys Leu Asn Gly Ala Asp Leu Lys Leu Arg Val Val Asp Gly Val Thr
260 265 270
Glu Cys Ser Gly Arg Leu Glu Val Lys Phe Gln Gly Glu Trp Gly Thr
275 280 285
Ile Cys Asp Asp Gly Trp Asp Ser Asp Asp Ala Ala Val Ala Cys Lys
290 295 300
Gln Leu Gly Cys Pro Thr Ala Val Thr Ala Ile Gly Arg Val Asn Ala
305 310 315 320
Ser Glu Gly Thr Gly His Ile Trp Leu Asp Ser Val Ser Cys His Gly
325 330 335
His Glu Ser Ala Leu Trp Gln Cys Arg His His Glu Trp Gly Lys His
340 345 350
Tyr Cys Asn His Asp Glu Asp Ala Gly Val Thr Cys Ser Asp Gly Ser
355 360 365
Asp Leu Glu Leu Arg Leu Lys Gly Gly Gly Ser His Cys Ala Gly Thr
370 375 380
Val Glu Val Glu Ile Gln Lys Leu Val Gly Lys Val Cys Asp Arg Ser
385 390 395 400
Trp Gly Leu Lys Glu Ala Asp Val Val Cys Arg Gln Leu Gly Cys Gly
405 410 415
Ser Ala Leu Lys Thr Ser Tyr Gln Val Tyr Ser Lys Thr Lys Ala Thr
420 425 430
Asn Thr Trp Leu Phe Val Ser Ser Cys Asn Gly Asn Glu Thr Ser Leu
435 440 445
Trp Asp Cys Lys Asn Trp Gln Trp Gly Gly Leu Ser Cys Asp His Tyr
450 455 460
Asp Glu Ala Lys Ile Thr Cys Ser Ala His Arg Lys Pro Arg Leu Val
465 470 475 480
Gly Gly Asp Ile Pro Cys Ser Gly Arg Val Glu Val Gln His Gly Asp
485 490 495
Thr Trp Gly Thr Val Cys Asp Ser Asp Phe Ser Leu Glu Ala Ala Ser
500 505 510
Val Leu Cys Arg Glu Leu Gln Cys Gly Thr Val Val Ser Leu Leu Gly
515 520 525
Gly Ala His Phe Gly Glu Gly Ser Gly Gln Ile Trp Ala Glu Glu Phe
530 535 540
Gln Cys Glu Gly His Glu Ser His Leu Ser Leu Cys Pro Val Ala Pro
545 550 555 560
Arg Pro Asp Gly Thr Cys Ser His Ser Arg Asp Val Gly Val Val Cys
565 570 575
Ser Arg Tyr Thr Gln Ile Arg Leu Val Asn Gly Lys Thr Pro Cys Glu
580 585 590
Gly Arg Val Glu Leu Asn Ile Leu Gly Ser Trp Gly Ser Leu Cys Asn
595 600 605
Ser His Trp Asp Met Glu Asp Ala His Val Leu Cys Gln Gln Leu Lys
610 615 620
Cys Gly Val Ala Leu Ser Ile Pro Gly Gly Ala Pro Phe Gly Lys Gly
625 630 635 640
Ser Glu Gln Val Trp Arg His Met Phe His Cys Thr Gly Thr Glu Lys
645 650 655
His Met Gly Asp Cys Ser Val Thr Ala Leu Gly Ala Ser Leu Cys Ser
660 665 670
Ser Gly Gln Val Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln Thr
675 680 685
Leu Ser Pro Cys Asn Ser Ser Ser Ser Asp Pro Ser Ser Ser Ile Ile
690 695 700
Ser Glu Glu Asn Gly Val Ala Cys Ile Gly Ser Gly Gln Leu Arg Leu
705 710 715 720
Val Asp Gly Gly Gly Arg Cys Ala Gly Arg Val Glu Val Tyr His Glu
725 730 735
Gly Ser Trp Gly Thr Ile Cys Asp Asp Ser Trp Asp Leu Asn Asp Ala
740 745 750
His Val Val Cys Lys Gln Leu Ser Cys Gly Trp Ala Ile Asn Ala Thr
755 760 765
Gly Ser Ala His Phe Gly Glu Gly Thr Gly Pro Ile Trp Leu Asp Glu
770 775 780
Ile Asn Cys Asn Gly Lys Glu Ser His Ile Trp Gln Cys His Ser His
785 790 795 800
Gly Trp Gly Arg His Asn Cys Arg His Lys Glu Asp Ala Gly Val Ile
805 810 815
Cys Ser Glu Phe Met Ser Leu Arg Leu Ile Ser Glu Asn Ser Arg Glu
820 825 830
Thr Cys Ala Gly Arg Leu Glu Val Phe Tyr Asn Gly Ala Trp Gly Ser
835 840 845
Val Gly Arg Asn Ser Met Ser Pro Ala Thr Val Gly Val Val Cys Arg
850 855 860
Gln Leu Gly Cys Ala Asp Arg Gly Asp Ile Ser Pro Ala Ser Ser Asp
865 870 875 880
Lys Thr Val Ser Arg His Met Trp Val Asp Asn Val Gln Cys Pro Lys
885 890 895
Gly Pro Asp Thr Leu Trp Gln Cys Pro Ser Ser Pro Trp Lys Lys Arg
900 905 910
Leu Ala Ser Pro Ser Glu Glu Thr Trp Ile Thr Cys Ala Asn Lys Ile
915 920 925
Arg Leu Gln Glu Gly Asn Thr Asn Cys Ser Gly Arg Val Glu Ile Trp
930 935 940
Tyr Gly Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu Glu
945 950 955 960
Asp Ala Gln Val Val Cys Arg Gln Leu Gly Cys Gly Ser Ala Leu Glu
965 970 975
Ala Gly Lys Glu Ala Ala Phe Gly Gln Gly Thr Gly Pro Ile Trp Leu
980 985 990
Asn Glu Val Lys Cys Lys Gly Asn Glu Thr Ser Leu Trp Asp Cys Pro
995 1000 1005
Ala Arg Ser Trp Gly His Ser Asp Cys Gly His Lys Glu Asp Ala
1010 1015 1020
Ala Val Thr Cys Ser Glu Ile Ala Lys Ser Arg Glu Ser Leu His
1025 1030 1035
Ala Thr Gly Arg Ser Ser Phe Val Ala Leu Ala Ile Phe Gly Val
1040 1045 1050
Ile Leu Leu Ala Cys Leu Ile Ala Phe Leu Ile Trp Thr Gln Lys
1055 1060 1065
Arg Arg Gln Arg Gln Arg Leu Ser Val Phe Ser Gly Gly Glu Asn
1070 1075 1080
Ser Val His Gln Ile Gln Tyr Arg Glu Met Asn Ser Cys Leu Lys
1085 1090 1095
Ala Asp Glu Thr Asp Met Leu Asn Pro Ser Gly Asp His Ser Glu
1100 1105 1110
Val Gln
1115
<210> SEQ ID NO 3
<211> LENGTH: 4405
<212> TYPE: DNA
<213> ORGANISM: mus musculus
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/AF274883
<309> DATABASE ENTRY DATE: 2001-05-10
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(4405)
<400> SEQUENCE: 3
gtggtcatcc actttctaca gagaacacgt ctatgaaata gtatcaggag acacacggag 60
ccatcaaaat catcaagctt tggaatgggt ggacacagaa tggttcttct tggaggtgct 120
ggatctcctg gttgtaaaag gtttgtccat ctaggtttct ttgttgtggc tgtgagctca 180
cttctcagtg cctctgctgt cactaacgct cctggagaaa tgaagaagga actgagactg 240
gcgggtggtg aaaacaactg tagtgggaga gtggaactta agatccatga caagtggggc 300
acagtgtgca gtaacggctg gagcatgaat gaagtgtccg tggtttgcca gcagctggga 360
tgcccaactt ctattaaagc ccttggatgg gctaactcca gcgccggctc tggatatatc 420
tggatggaca aagtttcttg tacagggaat gagtcagctc tttgggactg caaacatgat 480
gggtggggaa agcataactg tacccatgaa aaagatgctg gagtgacctg ctcagatgga 540
tctaatttgg agatgagact ggtgaacagt gcgggccacc gatgcttagg aagagtagaa 600
ataaagttcc agggaaagtg ggggacggtg tgtgacgaca acttcagcaa agatcacgct 660
tctgtgattt gtaaacagct tggatgtgga agtgccatta gtttctctgg ctcagctaaa 720
ttgggagctg gttctggacc aatctggctc gatgacctgg catgcaatgg aaatgagtca 780
gctctctggg actgcaaaca ccggggatgg ggcaagcata actgtgacca tgctgaggat 840
gtcggtgtga tttgcttaga gggagcagat ctgagcctga gactagtgga tggagtgtcc 900
agatgttcag gaagattgga agtgagattc caaggagaat gggggaccgt gtgtgatgat 960
aactgggatc tccgggatgc ttctgtggtg tgcaagcaac tgggatgtcc aactgccatc 1020
agtgccattg gtcgagttaa tgccagtgag ggatctggac agatttggct tgacaacatt 1080
tcatgcgaag gacatgaggc aactctttgg gagtgtaaac accaagagtg gggaaagcat 1140
tactgtcatc atagagaaga cgctggtgtg acatgttctg atggagcaga tctggaactt 1200
agacttgtag gtggaggcag tcgctgtgct ggcattgtgg aggtggagat tcagaagctg 1260
actgggaaga tgtgtagccg aggctggaca ctggcagatg cggatgtggt ttgcagacag 1320
cttggatgtg gatctgcgct tcaaacccag gctaagatct actctaaaac tggggcaaca 1380
aatacgtggc tctttcctgg atcttgtaat ggaaatgaaa ctactttttg gcaatgcaaa 1440
aactggcagt ggggcggcct ttcctgtgat aatttcgaag aagccaaagt tacctgctca 1500
ggccacaggg aacccagact ggttggagga gaaatcccat gctctggtcg tgtggaagtg 1560
aaacacggag acgtgtgggg ctccgtctgt gattttgact tgtctctgga agctgccagt 1620
gtggtgtgca gggaattaca atgtggaaca gtcgtctcta tcctaggggg agcacatttt 1680
ggagaaggaa gtggacagat ctggggtgaa gaattccagt gtagtgggga tgagtcccat 1740
ctttcactat gctcagtggc gcccccgcta gacagaactt gtacccacag cagggatgtc 1800
agcgtagtct gctcacgata catagatatt cgtctggcag gcggcgagtc ctcctgtgag 1860
ggaagagtgg agctcaagac actcggagcc tggggtcccc tctgcagttc tcattgggac 1920
atggaagatg ctcatgtctt atgtcagcag ctgaagtgtg gggttgccca atctattcca 1980
gaaggagcac attttgggaa aggagctggt caggtctgga gtcacatgtt ccactgcact 2040
ggaactgagg aacatatagg agattgcctc atgactgctc tgggtgcgcc gacgtgttcc 2100
gaaggacagg tggcctctgt catctgctca ggaaaccaat cccagacact attgccatgt 2160
agttcattgt ctccagtcca aacaacaagc tctacaattc caaaggagag tgaagttccc 2220
tgcatagcaa gtggccagct tcgcttggta ggtggaggtg gtcgctgcgc tggaagagtg 2280
gaggtctgcc acgagggctc ttggggcacc gtctgtgatg acaattggga tatgactgat 2340
gccaatgtgg tgtgcaagca gctggactgt ggcgtggcaa ttaacgccac tggctctgct 2400
tacttcgggg aaggagcagg agctatctgg ctagacgaag tcatctgcac tgggaaagag 2460
tctcatattt ggcagtgcca ttcacatggc tggggacgcc ataactgcag gcacaaagaa 2520
gatgcaggtg ttatctgctc cgagttcatg tctctgaggc tgaccaacga agcccacaaa 2580
gaaaactgca caggtcgcct tgaagtgttt tacaatggta catggggcag tattggcagt 2640
agcaatatgt ctccaaccac tgtgggggtg gtgtgccgtc agctgggctg tgcagacaac 2700
gggactgtga aacccatacc ttcagacaag acaccatcca ggcccatgtg ggtagatcgt 2760
gtgcagtgtc caaaaggagt tgacactttg tggcagtgcc cctcgtcacc ttggaaacag 2820
agacaggcca gcccctcctc ccaggagtcc tggatcatct gtgacaacaa aataagactc 2880
caggaagggc atacagactg ttctggacgt gtggagatct ggcacaaagg ttcctgggga 2940
acagtgtgtg atgactcctg ggatcttaat gatgctaagg ttgtatgtaa gcagttgggc 3000
tgtggccaag ctgtgaaggc actaaaagaa gcagcatttg gtccaggaac tgggcccata 3060
tggctcaatg aaattaagtg tagagggaat gagtcttccc tgtgggattg tcctgccaaa 3120
ccgtggagtc acagcgactg tgggcacaaa gaagatgctt ccatccagtg cctcccaaaa 3180
atgacttcag aatcacatca tggcacaggt caccccaccc tcacggcact cttggtttgt 3240
ggagccattc tattggtcct cctcattgtc ttcctcctgt ggactctgaa gcgacgacgg 3300
attcagcgac ttacagtttc ctcaagagga gaggtcttga tacatcaagt tcagtaccaa 3360
gagatggatt caaaggcgga tgatctggac ttgctgaaat cctcgggggt cattcagagg 3420
cacactgaga aggaaaatga taatttataa tccactgagg ttggagttta agaagccttg 3480
acaggacagc cagctaaatg gaacaagagc ccaggcaacg cacggatgac cacagctgca 3540
tcttcatgca gtcctttgtt tcctggaact ctgctgaacc tgcaaaaacc atatttgtga 3600
atgtgaccac ttaatagaga tgggagactt ttgagggaat taaacaatat tgctattggt 3660
ttgcttgttc gcaatagggt ctcattatgt atagccctgg agatggcgat agagagcagg 3720
caagcctaga attcacagag atctgcttgt ctctgcttcc caaatgctgg gatcaaatat 3780
gtggaccacc acatgtggtt taacaattgt gtcttgattt tataaatttc tggttggttt 3840
ttctgacgtt tttagggttt cgtgaatata aaataatgtc ttttcggttg gcatgctaat 3900
ttttaaatat tatacacttc cttgtagtga gtttaagaat aatttcttat aaccaagtca 3960
attcattttc actttgaatt atttaataaa ggaatatggt cattgtgacc acacacacag 4020
cagttgtgac cgcctgtatg aggccttcaa aaaatatttt aaaaatagag ggctggagaa 4080
atggctcaga ggtcctgagt tcaattccta gcaaccacat ggtggctcac aaccatctgt 4140
aatgggaatc cgatgccctc ttccagtgta tctgaagata gtgacactgt cctcattaac 4200
ataaaataaa taaataaatc tttaaaaaaa gaaaagacaa tagaggaggg gaggggcgtg 4260
agcgtaggag tgaggactga ttgagaagaa ggttggagga agtgggggga ggtgaaaagc 4320
tagttgggaa cttatgtgat cacagtgcat catgtccaaa tatgacatgt ccaaaaatgt 4380
tattaataaa gaaacggaaa tcaaa 4405
<210> SEQ ID NO 4
<211> LENGTH: 1121
<212> TYPE: PRT
<213> ORGANISM: Mus musculus
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/AAK16065
<309> DATABASE ENTRY DATE: 2001-05-10
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1121)
<400> SEQUENCE: 4
Met Gly Gly His Arg Met Val Leu Leu Gly Gly Ala Gly Ser Pro Gly
1 5 10 15
Cys Lys Arg Phe Val His Leu Gly Phe Phe Val Val Ala Val Ser Ser
20 25 30
Leu Leu Ser Ala Ser Ala Val Thr Asn Ala Pro Gly Glu Met Lys Lys
35 40 45
Glu Leu Arg Leu Ala Gly Gly Glu Asn Asn Cys Ser Gly Arg Val Glu
50 55 60
Leu Lys Ile His Asp Lys Trp Gly Thr Val Cys Ser Asn Gly Trp Ser
65 70 75 80
Met Asn Glu Val Ser Val Val Cys Gln Gln Leu Gly Cys Pro Thr Ser
85 90 95
Ile Lys Ala Leu Gly Trp Ala Asn Ser Ser Ala Gly Ser Gly Tyr Ile
100 105 110
Trp Met Asp Lys Val Ser Cys Thr Gly Asn Glu Ser Ala Leu Trp Asp
115 120 125
Cys Lys His Asp Gly Trp Gly Lys His Asn Cys Thr His Glu Lys Asp
130 135 140
Ala Gly Val Thr Cys Ser Asp Gly Ser Asn Leu Glu Met Arg Leu Val
145 150 155 160
Asn Ser Ala Gly His Arg Cys Leu Gly Arg Val Glu Ile Lys Phe Gln
165 170 175
Gly Lys Trp Gly Thr Val Cys Asp Asp Asn Phe Ser Lys Asp His Ala
180 185 190
Ser Val Ile Cys Lys Gln Leu Gly Cys Gly Ser Ala Ile Ser Phe Ser
195 200 205
Gly Ser Ala Lys Leu Gly Ala Gly Ser Gly Pro Ile Trp Leu Asp Asp
210 215 220
Leu Ala Cys Asn Gly Asn Glu Ser Ala Leu Trp Asp Cys Lys His Arg
225 230 235 240
Gly Trp Gly Lys His Asn Cys Asp His Ala Glu Asp Val Gly Val Ile
245 250 255
Cys Leu Glu Gly Ala Asp Leu Ser Leu Arg Leu Val Asp Gly Val Ser
260 265 270
Arg Cys Ser Gly Arg Leu Glu Val Arg Phe Gln Gly Glu Trp Gly Thr
275 280 285
Val Cys Asp Asp Asn Trp Asp Leu Arg Asp Ala Ser Val Val Cys Lys
290 295 300
Gln Leu Gly Cys Pro Thr Ala Ile Ser Ala Ile Gly Arg Val Asn Ala
305 310 315 320
Ser Glu Gly Ser Gly Gln Ile Trp Leu Asp Asn Ile Ser Cys Glu Gly
325 330 335
His Glu Ala Thr Leu Trp Glu Cys Lys His Gln Glu Trp Gly Lys His
340 345 350
Tyr Cys His His Arg Glu Asp Ala Gly Val Thr Cys Ser Asp Gly Ala
355 360 365
Asp Leu Glu Leu Arg Leu Val Gly Gly Gly Ser Arg Cys Ala Gly Ile
370 375 380
Val Glu Val Glu Ile Gln Lys Leu Thr Gly Lys Met Cys Ser Arg Gly
385 390 395 400
Trp Thr Leu Ala Asp Ala Asp Val Val Cys Arg Gln Leu Gly Cys Gly
405 410 415
Ser Ala Leu Gln Thr Gln Ala Lys Ile Tyr Ser Lys Thr Gly Ala Thr
420 425 430
Asn Thr Trp Leu Phe Pro Gly Ser Cys Asn Gly Asn Glu Thr Thr Phe
435 440 445
Trp Gln Cys Lys Asn Trp Gln Trp Gly Gly Leu Ser Cys Asp Asn Phe
450 455 460
Glu Glu Ala Lys Val Thr Cys Ser Gly His Arg Glu Pro Arg Leu Val
465 470 475 480
Gly Gly Glu Ile Pro Cys Ser Gly Arg Val Glu Val Lys His Gly Asp
485 490 495
Val Trp Gly Ser Val Cys Asp Phe Asp Leu Ser Leu Glu Ala Ala Ser
500 505 510
Val Val Cys Arg Glu Leu Gln Cys Gly Thr Val Val Ser Ile Leu Gly
515 520 525
Gly Ala His Phe Gly Glu Gly Ser Gly Gln Ile Trp Gly Glu Glu Phe
530 535 540
Gln Cys Ser Gly Asp Glu Ser His Leu Ser Leu Cys Ser Val Ala Pro
545 550 555 560
Pro Leu Asp Arg Thr Cys Thr His Ser Arg Asp Val Ser Val Val Cys
565 570 575
Ser Arg Tyr Ile Asp Ile Arg Leu Ala Gly Gly Glu Ser Ser Cys Glu
580 585 590
Gly Arg Val Glu Leu Lys Thr Leu Gly Ala Trp Gly Pro Leu Cys Ser
595 600 605
Ser His Trp Asp Met Glu Asp Ala His Val Leu Cys Gln Gln Leu Lys
610 615 620
Cys Gly Val Ala Gln Ser Ile Pro Glu Gly Ala His Phe Gly Lys Gly
625 630 635 640
Ala Gly Gln Val Trp Ser His Met Phe His Cys Thr Gly Thr Glu Glu
645 650 655
His Ile Gly Asp Cys Leu Met Thr Ala Leu Gly Ala Pro Thr Cys Ser
660 665 670
Glu Gly Gln Val Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln Thr
675 680 685
Leu Leu Pro Cys Ser Ser Leu Ser Pro Val Gln Thr Thr Ser Ser Thr
690 695 700
Ile Pro Lys Glu Ser Glu Val Pro Cys Ile Ala Ser Gly Gln Leu Arg
705 710 715 720
Leu Val Gly Gly Gly Gly Arg Cys Ala Gly Arg Val Glu Val Tyr His
725 730 735
Glu Gly Ser Trp Gly Thr Val Cys Asp Asp Asn Trp Asp Met Thr Asp
740 745 750
Ala Asn Val Val Cys Lys Gln Leu Asp Cys Gly Val Ala Ile Asn Ala
755 760 765
Thr Gly Ser Ala Tyr Phe Gly Glu Gly Ala Gly Ala Ile Trp Leu Asp
770 775 780
Glu Val Ile Cys Thr Gly Lys Glu Ser His Ile Trp Gln Cys His Ser
785 790 795 800
His Gly Trp Gly Arg His Asn Cys Arg His Lys Glu Asp Ala Gly Val
805 810 815
Ile Cys Ser Glu Phe Met Ser Leu Arg Leu Thr Asn Glu Ala His Lys
820 825 830
Glu Ser Cys Thr Gly Arg Leu Glu Val Phe Tyr Asn Gly Thr Trp Gly
835 840 845
Ser Ile Gly Ser Ser Asn Met Ser Pro Thr Thr Val Gly Val Val Cys
850 855 860
Arg Gln Leu Gly Cys Ala Asp Asn Gly Thr Val Lys Pro Ile Pro Ser
865 870 875 880
Asp Lys Thr Pro Ser Arg Pro Met Trp Val Asp Arg Val Gln Cys Pro
885 890 895
Lys Gly Val Asp Thr Leu Trp Gln Cys Pro Ser Ser Pro Trp Lys Gln
900 905 910
Arg Gln Ala Ser Pro Ser Ser Gln Glu Ser Trp Ile Ile Cys Asp Asn
915 920 925
Lys Ile Arg Leu Gln Glu Gly His Thr Asp Cys Ser Gly Arg Val Glu
930 935 940
Ile Trp His Lys Gly Phe Trp Gly Thr Val Cys Asp Asp Ser Trp Asp
945 950 955 960
Leu Asn Asp Ala Lys Val Val Cys Lys Gln Leu Gly Cys Gly Gln Ala
965 970 975
Val Lys Ala Leu Lys Glu Ala Ala Phe Gly Pro Gly Thr Gly Pro Ile
980 985 990
Trp Leu Asn Glu Ile Lys Cys Arg Gly Asn Glu Ser Ser Leu Trp Asp
995 1000 1005
Cys Pro Ala Lys Pro Trp Ser His Ser Asp Cys Gly His Lys Glu
1010 1015 1020
Asp Ala Ser Ile Gln Cys Leu Pro Lys Met Thr Ser Glu Ser His
1025 1030 1035
His Gly Thr Gly His Pro Thr Leu Thr Ala Leu Leu Val Cys Gly
1040 1045 1050
Ala Ile Leu Leu Val Leu Leu Ile Val Phe Leu Leu Trp Thr Leu
1055 1060 1065
Lys Arg Arg Gln Ile Gln Arg Leu Thr Val Ser Ser Arg Gly Glu
1070 1075 1080
Val Leu Ile His Gln Val Gln Tyr Gln Glu Met Asp Ser Lys Ala
1085 1090 1095
Asp Asp Leu Asp Leu Leu Lys Ser Ser Gly Val Ile Gln Arg His
1100 1105 1110
Thr Glu Lys Glu Asn Asp Asn Leu
1115 1120
<210> SEQ ID NO 5
<211> LENGTH: 3780
<212> TYPE: DNA
<213> ORGANISM: homo sapiens
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/BC051281
<309> DATABASE ENTRY DATE: 2006-07-15
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(3780)
<400> SEQUENCE: 5
aacatttcta gggaataata caagaagatt taggaatcat tgaagttata aatctttgga 60
atgagcaaac tcagaatggt gctacttgaa gactctggat ctgctgactt cagaagacat 120
tttgtcaact tgagtccctt caccattact gtggtcttac ttctcagtgc ctgttttgtc 180
accagttctc ttggaggaac agacaaggag ctgaggctag tggatggtga aaacaagtgt 240
agcgggagag tggaagtgaa agtccaggag gagtggggaa cggtgtgtaa taatggctgg 300
agcatggaag cggtctctgt gatttgtaac cagctgggat gtccaactgc tatcaaagcc 360
cctggatggg ctaattccag tgcaggttct ggacgcattt ggatggatca tgtttcttgt 420
cgtgggaatg agtcagctct ttgggattgc aaacatgatg gatggggaaa gcatagtaac 480
tgtactcacc aacaagatgc tggagtgacc tgctcagatg gatccaattt ggaaatgagg 540
ctgacgcgtg gagggaatat gtgttctgga agaatagaga tcaaattcca aggacggtgg 600
ggaacagtgt gtgatgataa cttcaacata gatcatgcat ctgtcatttg tagacaactt 660
gaatgtggaa gtgctgtcag tttctctggt tcatctaatt ttggagaagg ctctggacca 720
atctggtttg atgatcttat atgcaacgga aatgagtcag ctctctggaa ctgcaaacat 780
caaggatggg gaaagcataa ctgtgatcat gctgaggatg ctggagtgat ttgctcaaag 840
ggagcagatc tgagcctgag actggtagat ggagtcactg aatgttcagg aagattagaa 900
gtgagattcc aaggagaatg ggggacaata tgtgatgacg gctgggacag ttacgatgct 960
gctgtggcat gcaagcaact gggatgtcca actgccgtca cagccattgg tcgagttaac 1020
gccagtaagg gatttggaca catctggctt gacagcgttt cttgccaggg acatgaacct 1080
gctgtctggc aatgtaaaca ccatgaatgg ggaaagcatt attgcaatca caatgaagat 1140
gctggcgtga catgttctga tggatcagat ctggagctaa gacttagagg tggaggcagc 1200
cgctgtgctg ggacagttga ggtggagatt cagagactgt tagggaaggt gtgtgacaga 1260
ggctggggac tgaaagaagc tgatgtggtt tgcaggcagc tgggatgtgg atctgcactc 1320
aaaacatctt atcaagtgta ctccaaaatc caggcaacaa acacatggct gtttctaagt 1380
agctgtaacg gaaatgaaac ttctctttgg gactgcaaga actggcaatg gggtggactt 1440
acctgtgatc actatgaaga agccaaaatt acctgctcag cccacaggga acccagactg 1500
gttggagggg acattccctg ttctggacgt gttgaagtga agcatggtga cacgtggggc 1560
tccatctgtg attcggactt ctctctggaa gctgccagcg ttctatgcag ggaattacag 1620
tgtggcacag ttgtctctat cctgggggga gctcactttg gagagggaaa tggacagatc 1680
tgggctgaag aattccagtg tgagggacat gagtcccatc tttcactctg cccagtagca 1740
ccccgcccag aaggaacttg tagccacagc agggatgttg gagtagtctg ctcaagatac 1800
acagaaattc gcttggtgaa tggcaagacc ccgtgtgagg gcagagtgga gctcaaaacg 1860
cttggtgcct ggggatccct ctgtaactct cactgggaca tagaagatgc ccatgttctt 1920
tgccagcagc ttaaatgtgg agttgccctt tctaccccag gaggagcacg ttttggaaaa 1980
ggaaatggtc agatctggag gcatatgttt cactgcactg ggactgagca gcacatggga 2040
gattgtcctg taactgctct aggtgcttca ttatgtcctt cagagcaagt ggcctctgta 2100
atctgctcag gaaaccagtc ccaaacactg tcctcgtgca attcatcgtc tttgggccca 2160
acaaggccta ccattccaga agaaagtgct gtggcctgca tagagagtgg tcaacttcgc 2220
ctggtaaatg gaggaggtcg ctgtgctggg agagtagaga tctatcatga gggctcctgg 2280
ggcaccatct gtgatgacag ctgggacctg agtgatgccc acgtggtttg cagacagctg 2340
ggctgtggag aggccattaa tgccactggt tctgctcatt ttggggaagg aacagggccc 2400
atctggctgg atgagatgaa atgcaatgga aaagaatccc gcatttggca gtgccattca 2460
cacggctggg ggcagcaaaa ttgcaggcac aaggaggatg cgggagttat ctgctcagaa 2520
ttcatgtctc tgagactgac cagtgaagcc agcagagagg cctgtgcagg gcgtctggaa 2580
gttttttaca atggagcttg gggcactgtt ggcaagagta gcatgtctga aaccactgtg 2640
ggtgtggtgt gcaggcagct gggctgtgca gacaaaggga aaatcaaccc tgcatcttta 2700
gacaaggcca tgtccattcc catgtgggtg gacaatgttc agtgtccaaa aggacctgac 2760
acgctgtggc agtgcccatc atctccatgg gagaagagac tggccagccc ctcggaggag 2820
acctggatca catgtgacaa caagataaga cttcaggaag gacccacttc ctgttctgga 2880
cgtgtggaga tctggcatgg aggttcctgg gggacagtgt gtgatgactc ttgggacttg 2940
gacgatgctc aggtggtgtg tcaacaactt ggctgtggtc cagctttgaa agcattcaaa 3000
gaagcagagt ttggtcaggg gactggaccg atatggctca atgaagtgaa gtgcaaaggg 3060
aatgagtctt ccttgtggga ttgtcctgcc agacgctggg gccatagtga gtgtgggcac 3120
aaggaagacg ctgcagtgaa ttgcacagat atttcagtgc agaaaacccc acaaaaagcc 3180
acaacaggtc gctcatcccg tcagtcatcc tttattgcag tcgggatcct tggggttgtt 3240
ctgttggcca ttttcgtcgc attattcttc ttgactaaaa agcgaagaca gagacagcgg 3300
cttgcagttt cctcaagagg agagaactta gtccaccaaa ttcaataccg ggagatgaat 3360
tcttgcctga atgcagatga tctggaccta atgaattcct cagaaaattc ccatgagtca 3420
gctgatttca gtgctgctga actaatttct gtgtctaaat ttcttcctat ttctggaatg 3480
gaaaaggagg ccattctgag ccacactgaa aaggaaaatg ggaatttata acccagtgag 3540
ttcagccttt aagatacctt gatgaagacc tggactattg aatggagcag aaattcacct 3600
ctctcactga ctattacagt tgcattttta tggagttctt cttctcctag gattcctaag 3660
actgctgctg aatttataaa aattaagttt gtgaatgtga ctacttagtg gtgtatatga 3720
gactttcaag ggaattaaat aaataaataa gaatgttatt gaaaaaaaaa aaaaaaaaaa 3780
<210> SEQ ID NO 6
<211> LENGTH: 1156
<212> TYPE: PRT
<213> ORGANISM: HOMO SAPIENS
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/AAH51281
<309> DATABASE ENTRY DATE: 2006-07-15
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1156)
<400> SEQUENCE: 6
Met Ser Lys Leu Arg Met Val Leu Leu Glu Asp Ser Gly Ser Ala Asp
1 5 10 15
Phe Arg Arg His Phe Val Asn Leu Ser Pro Phe Thr Ile Thr Val Val
20 25 30
Leu Leu Leu Ser Ala Cys Phe Val Thr Ser Ser Leu Gly Gly Thr Asp
35 40 45
Lys Glu Leu Arg Leu Val Asp Gly Glu Asn Lys Cys Ser Gly Arg Val
50 55 60
Glu Val Lys Val Gln Glu Glu Trp Gly Thr Val Cys Asn Asn Gly Trp
65 70 75 80
Ser Met Glu Ala Val Ser Val Ile Cys Asn Gln Leu Gly Cys Pro Thr
85 90 95
Ala Ile Lys Ala Pro Gly Trp Ala Asn Ser Ser Ala Gly Ser Gly Arg
100 105 110
Ile Trp Met Asp His Val Ser Cys Arg Gly Asn Glu Ser Ala Leu Trp
115 120 125
Asp Cys Lys His Asp Gly Trp Gly Lys His Ser Asn Cys Thr His Gln
130 135 140
Gln Asp Ala Gly Val Thr Cys Ser Asp Gly Ser Asn Leu Glu Met Arg
145 150 155 160
Leu Thr Arg Gly Gly Asn Met Cys Ser Gly Arg Ile Glu Ile Lys Phe
165 170 175
Gln Gly Arg Trp Gly Thr Val Cys Asp Asp Asn Phe Asn Ile Asp His
180 185 190
Ala Ser Val Ile Cys Arg Gln Leu Glu Cys Gly Ser Ala Val Ser Phe
195 200 205
Ser Gly Ser Ser Asn Phe Gly Glu Gly Ser Gly Pro Ile Trp Phe Asp
210 215 220
Asp Leu Ile Cys Asn Gly Asn Glu Ser Ala Leu Trp Asn Cys Lys His
225 230 235 240
Gln Gly Trp Gly Lys His Asn Cys Asp His Ala Glu Asp Ala Gly Val
245 250 255
Ile Cys Ser Lys Gly Ala Asp Leu Ser Leu Arg Leu Val Asp Gly Val
260 265 270
Thr Glu Cys Ser Gly Arg Leu Glu Val Arg Phe Gln Gly Glu Trp Gly
275 280 285
Thr Ile Cys Asp Asp Gly Trp Asp Ser Tyr Asp Ala Ala Val Ala Cys
290 295 300
Lys Gln Leu Gly Cys Pro Thr Ala Val Thr Ala Ile Gly Arg Val Asn
305 310 315 320
Ala Ser Lys Gly Phe Gly His Ile Trp Leu Asp Ser Val Ser Cys Gln
325 330 335
Gly His Glu Pro Ala Val Trp Gln Cys Lys His His Glu Trp Gly Lys
340 345 350
His Tyr Cys Asn His Asn Glu Asp Ala Gly Val Thr Cys Ser Asp Gly
355 360 365
Ser Asp Leu Glu Leu Arg Leu Arg Gly Gly Gly Ser Arg Cys Ala Gly
370 375 380
Thr Val Glu Val Glu Ile Gln Arg Leu Leu Gly Lys Val Cys Asp Arg
385 390 395 400
Gly Trp Gly Leu Lys Glu Ala Asp Val Val Cys Arg Gln Leu Gly Cys
405 410 415
Gly Ser Ala Leu Lys Thr Ser Tyr Gln Val Tyr Ser Lys Ile Gln Ala
420 425 430
Thr Asn Thr Trp Leu Phe Leu Ser Ser Cys Asn Gly Asn Glu Thr Ser
435 440 445
Leu Trp Asp Cys Lys Asn Trp Gln Trp Gly Gly Leu Thr Cys Asp His
450 455 460
Tyr Glu Glu Ala Lys Ile Thr Cys Ser Ala His Arg Glu Pro Arg Leu
465 470 475 480
Val Gly Gly Asp Ile Pro Cys Ser Gly Arg Val Glu Val Lys His Gly
485 490 495
Asp Thr Trp Gly Ser Ile Cys Asp Ser Asp Phe Ser Leu Glu Ala Ala
500 505 510
Ser Val Leu Cys Arg Glu Leu Gln Cys Gly Thr Val Val Ser Ile Leu
515 520 525
Gly Gly Ala His Phe Gly Glu Gly Asn Gly Gln Ile Trp Ala Glu Glu
530 535 540
Phe Gln Cys Glu Gly His Glu Ser His Leu Ser Leu Cys Pro Val Ala
545 550 555 560
Pro Arg Pro Glu Gly Thr Cys Ser His Ser Arg Asp Val Gly Val Val
565 570 575
Cys Ser Arg Tyr Thr Glu Ile Arg Leu Val Asn Gly Lys Thr Pro Cys
580 585 590
Glu Gly Arg Val Glu Leu Lys Thr Leu Gly Ala Trp Gly Ser Leu Cys
595 600 605
Asn Ser His Trp Asp Ile Glu Asp Ala His Val Leu Cys Gln Gln Leu
610 615 620
Lys Cys Gly Val Ala Leu Ser Thr Pro Gly Gly Ala Arg Phe Gly Lys
625 630 635 640
Gly Asn Gly Gln Ile Trp Arg His Met Phe His Cys Thr Gly Thr Glu
645 650 655
Gln His Met Gly Asp Cys Pro Val Thr Ala Leu Gly Ala Ser Leu Cys
660 665 670
Pro Ser Glu Gln Val Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln
675 680 685
Thr Leu Ser Ser Cys Asn Ser Ser Ser Leu Gly Pro Thr Arg Pro Thr
690 695 700
Ile Pro Glu Glu Ser Ala Val Ala Cys Ile Glu Ser Gly Gln Leu Arg
705 710 715 720
Leu Val Asn Gly Gly Gly Arg Cys Ala Gly Arg Val Glu Ile Tyr His
725 730 735
Glu Gly Ser Trp Gly Thr Ile Cys Asp Asp Ser Trp Asp Leu Ser Asp
740 745 750
Ala His Val Val Cys Arg Gln Leu Gly Cys Gly Glu Ala Ile Asn Ala
755 760 765
Thr Gly Ser Ala His Phe Gly Glu Gly Thr Gly Pro Ile Trp Leu Asp
770 775 780
Glu Met Lys Cys Asn Gly Lys Glu Ser Arg Ile Trp Gln Cys His Ser
785 790 795 800
His Gly Trp Gly Gln Gln Asn Cys Arg His Lys Glu Asp Ala Gly Val
805 810 815
Ile Cys Ser Glu Phe Met Ser Leu Arg Leu Thr Ser Glu Ala Ser Arg
820 825 830
Glu Ala Cys Ala Gly Arg Leu Glu Val Phe Tyr Asn Gly Ala Trp Gly
835 840 845
Thr Val Gly Lys Ser Ser Met Ser Glu Thr Thr Val Gly Val Val Cys
850 855 860
Arg Gln Leu Gly Cys Ala Asp Lys Gly Lys Ile Asn Pro Ala Ser Leu
865 870 875 880
Asp Lys Ala Met Ser Ile Pro Met Trp Val Asp Asn Val Gln Cys Pro
885 890 895
Lys Gly Pro Asp Thr Leu Trp Gln Cys Pro Ser Ser Pro Trp Glu Lys
900 905 910
Arg Leu Ala Ser Pro Ser Glu Glu Thr Trp Ile Thr Cys Asp Asn Lys
915 920 925
Ile Arg Leu Gln Glu Gly Pro Thr Ser Cys Ser Gly Arg Val Glu Ile
930 935 940
Trp His Gly Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu
945 950 955 960
Asp Asp Ala Gln Val Val Cys Gln Gln Leu Gly Cys Gly Pro Ala Leu
965 970 975
Lys Ala Phe Lys Glu Ala Glu Phe Gly Gln Gly Thr Gly Pro Ile Trp
980 985 990
Leu Asn Glu Val Lys Cys Lys Gly Asn Glu Ser Ser Leu Trp Asp Cys
995 1000 1005
Pro Ala Arg Arg Trp Gly His Ser Glu Cys Gly His Lys Glu Asp
1010 1015 1020
Ala Ala Val Asn Cys Thr Asp Ile Ser Val Gln Lys Thr Pro Gln
1025 1030 1035
Lys Ala Thr Thr Gly Arg Ser Ser Arg Gln Ser Ser Phe Ile Ala
1040 1045 1050
Val Gly Ile Leu Gly Val Val Leu Leu Ala Ile Phe Val Ala Leu
1055 1060 1065
Phe Phe Leu Thr Lys Lys Arg Arg Gln Arg Gln Arg Leu Ala Val
1070 1075 1080
Ser Ser Arg Gly Glu Asn Leu Val His Gln Ile Gln Tyr Arg Glu
1085 1090 1095
Met Asn Ser Cys Leu Asn Ala Asp Asp Leu Asp Leu Met Asn Ser
1100 1105 1110
Ser Glu Asn Ser His Glu Ser Ala Asp Phe Ser Ala Ala Glu Leu
1115 1120 1125
Ile Ser Val Ser Lys Phe Leu Pro Ile Ser Gly Met Glu Lys Glu
1130 1135 1140
Ala Ile Leu Ser His Thr Glu Lys Glu Asn Gly Asn Leu
1145 1150 1155
<210> SEQ ID NO 7
<211> LENGTH: 4950
<212> TYPE: DNA
<213> ORGANISM: homo sapiens
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/Z22970
<309> DATABASE ENTRY DATE: 2005-04-18
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(4950)
<400> SEQUENCE: 7
gaattcttag ttgttttctt tagaagaaca tttctaggga ataatacaag aagatttagg 60
aatcattgaa gttataaatc tttggaatga gcaaactcag aatggtgcta cttgaagact 120
ctggatctgc tgacttcaga agacattttg tcaacctgag tcccttcacc attactgtgg 180
tcttacttct cagtgcctgt tttgtcacca gttctcttgg aggaacagac aaggagctga 240
ggctagtgga tggtgaaaac aagtgtagcg ggagagtgga agtgaaagtc caggaggagt 300
ggggaacggt gtgtaataat ggctggagca tggaagcggt ctctgtgatt tgtaaccagc 360
tgggatgtcc aactgctatc aaagcccctg gatgggctaa ttccagtgca ggttctggac 420
gcatttggat ggatcatgtt tcttgtcgtg ggaatgagtc agctctttgg gattgcaaac 480
atgatggatg gggaaagcat agtaactgta ctcaccaaca agatgctgga gtgacctgct 540
cagatggatc caatttggaa atgaggctga cgcgtggagg gaatatgtgt tctggaagaa 600
tagagatcaa attccaagga cggtggggaa cagtgtgtga tgataacttc aacatagatc 660
atgcatctgt catttgtaga caacttgaat gtggaagtgc tgtcagtttc tctggttcat 720
ctaattttgg agaaggctct ggaccaatct ggtttgatga tcttatatgc aacggaaatg 780
agtcagctct ctggaactgc aaacatcaag gatggggaaa gcataactgt gatcatgctg 840
aggatgctgg agtgatttgc tcaaagggag cagatctgag cctgagactg gtagatggag 900
tcactgaatg ttcaggaaga ttagaagtga gattccaagg agaatggggg acaatatgtg 960
atgacggctg ggacagttac gatgctgctg tggcatgcaa gcaactggga tgtccaactg 1020
ccgtcacagc cattggtcga gttaacgcca gtaagggatt tggacacatc tggcttgaca 1080
gcgtttcttg ccagggacat gaacctgctg tctggcaatg taaacaccat gaatggggaa 1140
agcattattg caatcacaat gaagatgctg gcgtgacatg ttctgatgga tcagatctgg 1200
agctaagact tagaggtgga ggcagccgct gtgctgggac agttgaggtg gagattcaga 1260
gactgttagg gaaggtgtgt gacagaggct ggggactgaa agaagctgat gtggtttgca 1320
ggcagctggg atgtggatct gcactcaaaa catcttatca agtgtactcc aaaatccagg 1380
caacaaacac atggctgttt ctaagtagct gtaacggaaa tgaaacttct ctttgggact 1440
gcaagaactg gcaatggggt ggacttacct gtgatcacta tgaagaagcc aaaattacct 1500
gctcagccca cagggaaccc agactggttg gaggggacat tccctgttct ggacgtgttg 1560
aagtgaagca tggtgacacg tggggctcca tctgtgattc ggacttctct ctggaagctg 1620
ccagcgttct atgcagggaa ttacagtgtg gcacagttgt ctctatcctg gggggagctc 1680
actttggaga gggaaatgga cagatctggg ctgaagaatt ccagtgtgag ggacatgagt 1740
cccatctttc actctgccca gtagcacccc gcccagaagg aacttgtagc cacagcaggg 1800
atgttggagt agtctgctca agatacacag aaattcgctt ggtgaatggc aagaccccgt 1860
gtgagggcag agtggagctc aaaacgcttg gtgcctgggg atccctctgt aactctcact 1920
gggacataga agatgcccat gttctttgcc agcagcttaa atgtggagtt gccctttcta 1980
ccccaggagg agcacgtttt ggaaaaggaa atggtcagat ctggaggcat atgtttcact 2040
gcactgggac tgagcagcac atgggagatt gtcctgtaac tgctctaggt gcttcattat 2100
gtccttcaga gcaagtggcc tctgtaatct gctcaggaaa ccagtcccaa acactgtcct 2160
cgtgcaattc atcgtctttg ggcccaacaa ggcctaccat tccagaagaa agtgctgtgg 2220
cctgcataga gagtggtcaa cttcgcctgg taaatggagg aggtcgctgt gctgggagag 2280
tagagatcta tcatgagggc tcctggggca ccatctgtga tgacagctgg gacctgagtg 2340
atgcccacgt ggtttgcaga cagctgggct gtggagaggc cattaatgcc actggttctg 2400
ctcattttgg ggaaggaaca gggcccatct ggctggatga gatgaaatgc aatggaaaag 2460
aatcccgcat ttggcagtgc cattcacacg gctgggggca gcaaaattgc aggcacaagg 2520
aggatgcggg agttatctgc tcagaattca tgtctctgag actgaccagt gaagccagca 2580
gagaggcctg tgcagggcgt ctggaagttt tttacaatgg agcttggggc actgttggca 2640
agagtagcat gtctgaaacc actgtgggtg tggtgtgcag gcagctgggc tgtgcagaca 2700
aagggaaaat caaccctgca tctttagaca aggccatgtc cattcccatg tgggtggaca 2760
atgttcagtg tccaaaagga cctgacacgc tgtggcagtg cccatcatct ccatgggaga 2820
agagactggc cagcccctcg gaggagacct ggatcacatg tgacaacaag ataagacttc 2880
aggaaggacc cacttcctgt tctggacgtg tggagatctg gcatggaggt tcctggggga 2940
cagtgtgtga tgactcttgg gacttggacg atgctcaggt ggtgtgtcaa caacttggct 3000
gtggtccagc tttgaaagca ttcaaagaag cagagtttgg tcaggggact ggaccgatat 3060
ggctcaatga agtgaagtgc aaagggaatg agtcttcctt gtgggattgt cctgccagac 3120
gctggggcca tagtgagtgt gggcacaagg aagacgctgc agtgaattgc acagatattt 3180
cagtgcagaa aaccccacaa aaagccacaa caggtcgctc atcccgtcag tcatccttta 3240
ttgcagtcgg gatccttggg gttgttctgt tggccatttt cgtcgcatta ttcttcttga 3300
ctaaaaagcg aagacagaga cagcggcttg cagtttcctc aagaggagag aacttagtcc 3360
accaaattca ataccgggag atgaattctt gcctgaatgc agatgatctg gacctaatga 3420
attcctcagg tctgtgggtt cttggagggt ctattgccca ggggttcaga tcagtggctg 3480
cagttgaggc acagacattc tactttgata aacagttaaa aaagtctaaa aatgtaatag 3540
gaagcttaga tgcatataat ggacaagaat gactgaaaat tattcttgga gaatatcaaa 3600
attgcaatca tagggaggcc tttagcttaa gaggcctgtg attattcctg atagaggtat 3660
ggaaagaacc atgcagagga atattatgac ttggacctca ttttattaaa acagaaatta 3720
atcttacaaa agattgtcat aagtgacagt ttaacttttt tctttaaatt ttgttgtgta 3780
tatttaaggt atacaacatg attttatggg atgtatatag atagtaaaaa gcttactaaa 3840
gcaaagcaaa tgaacacacc catcatctga catagttacc cttttttgtg ttgttcttgt 3900
ggcaagagca gctaaaacct actcacttag catgaatcct acatacagca caatgttatt 3960
acctataatc ctcatgttgt acattagacc tctagactgg ttcattctac gtatctgcta 4020
ctttgtatcc tctgacctac atacgtcttt cacagtttct tccattccca tttcctgtca 4080
ttttttttct ctagcttgat atttattata tttttcccta aaagtctaaa accttaaact 4140
ttcaatatct ttattgcatg agaagccata caaatccaca gaactagcct tatttctcat 4200
cacatcatgc tgttttatcc ttgaacttct atttagcacc agtgcactaa ttctgcatct 4260
gggcaggatg actttactgg gttggaagaa atatcccaaa acccattgtc tttactccat 4320
gaagggtccc tgaccttctg agaggggcct gcctcacttc ttccatccaa agaattatgc 4380
atctgctact gtgtcaggga acatatttaa ggaacatgta ctgttactgt gtcaggaaac 4440
atatttaaga aataggaaag actttctctg ccccttaaat cacacatgct tttcttccta 4500
gttatgggtg gtgtttttag ttgctcaaag agcctcacag ttacgtgaga agaggtctgg 4560
tttatttccc agtaattatt ttcttccttt cagaaaattc ccatgagtca gctgatttca 4620
gtgctgctga actaatttct gtgtctaaat ttcttcctat ttctggaatg gaaaaggagg 4680
ccattctgag ccacactgaa aaggaaaatg ggaatttata acccagtgag ttcagccttt 4740
aagatacctt gatgaagacc tggactattg aatggagcag aaattcacct ctctcactga 4800
ctattacagt tgcattttta tggagttctt cttctcctag gattcctaag actgctgctg 4860
aatttataaa aattaagttt gtgaatgtga ctacttagtg gtgtatatga gactttcaag 4920
ggaattaaat aaataaataa gaatgttaaa 4950
<210> SEQ ID NO 8
<211> LENGTH: 1156
<212> TYPE: PRT
<213> ORGANISM: HOMO SAPIENS
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/CAA80543
<309> DATABASE ENTRY DATE: 2005-04-18
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1156)
<400> SEQUENCE: 8
Met Val Leu Leu Glu Asp Ser Gly Ser Ala Asp Phe Arg Arg His Phe
1 5 10 15
Val Asn Leu Ser Pro Phe Thr Ile Thr Val Val Leu Leu Leu Ser Ala
20 25 30
Cys Phe Val Thr Ser Ser Leu Gly Gly Thr Asp Lys Glu Leu Arg Leu
35 40 45
Val Asp Gly Glu Asn Lys Cys Ser Gly Arg Val Glu Val Lys Val Gln
50 55 60
Glu Glu Trp Gly Thr Val Cys Asn Asn Gly Trp Ser Met Glu Ala Val
65 70 75 80
Ser Val Ile Cys Asn Gln Leu Gly Cys Pro Thr Ala Ile Lys Ala Pro
85 90 95
Gly Trp Ala Asn Ser Ser Ala Gly Ser Gly Arg Ile Trp Met Asp His
100 105 110
Val Ser Cys Arg Gly Asn Glu Ser Ala Leu Trp Asp Cys Lys His Asp
115 120 125
Gly Trp Gly Lys His Ser Asn Cys Thr His Gln Gln Asp Ala Gly Val
130 135 140
Thr Cys Ser Asp Gly Ser Asn Leu Glu Met Arg Leu Thr Arg Gly Gly
145 150 155 160
Asn Met Cys Ser Gly Arg Ile Glu Ile Lys Phe Gln Gly Arg Trp Gly
165 170 175
Thr Val Cys Asp Asp Asn Phe Asn Ile Asp His Ala Ser Val Ile Cys
180 185 190
Arg Gln Leu Glu Cys Gly Ser Ala Val Ser Phe Ser Gly Ser Ser Asn
195 200 205
Phe Gly Glu Gly Ser Gly Pro Ile Trp Phe Asp Asp Leu Ile Cys Asn
210 215 220
Gly Asn Glu Ser Ala Leu Trp Asn Cys Lys His Gln Gly Trp Gly Lys
225 230 235 240
His Asn Cys Asp His Ala Glu Asp Ala Gly Val Ile Cys Ser Lys Gly
245 250 255
Ala Asp Leu Ser Leu Arg Leu Val Asp Gly Val Thr Glu Cys Ser Gly
260 265 270
Arg Leu Glu Val Arg Phe Gln Gly Glu Trp Gly Thr Ile Cys Asp Asp
275 280 285
Gly Trp Asp Ser Tyr Asp Ala Ala Val Ala Cys Lys Gln Leu Gly Cys
290 295 300
Pro Thr Ala Val Thr Ala Ile Gly Arg Val Asn Ala Ser Lys Gly Phe
305 310 315 320
Gly His Ile Trp Leu Asp Ser Val Ser Cys Gln Gly His Glu Pro Ala
325 330 335
Val Trp Gln Cys Lys His His Glu Trp Gly Lys His Tyr Cys Asn His
340 345 350
Asn Glu Asp Ala Gly Val Thr Cys Ser Asp Gly Ser Asp Leu Glu Leu
355 360 365
Arg Leu Arg Gly Gly Gly Ser Arg Cys Ala Gly Thr Val Glu Val Glu
370 375 380
Ile Gln Arg Leu Leu Gly Lys Val Cys Asp Arg Gly Trp Gly Leu Lys
385 390 395 400
Glu Ala Asp Val Val Cys Arg Gln Leu Gly Cys Gly Ser Ala Leu Lys
405 410 415
Thr Ser Tyr Gln Val Tyr Ser Lys Ile Gln Ala Thr Asn Thr Trp Leu
420 425 430
Phe Leu Ser Ser Cys Asn Gly Asn Glu Thr Ser Leu Trp Asp Cys Lys
435 440 445
Asn Trp Gln Trp Gly Gly Leu Thr Cys Asp His Tyr Glu Glu Ala Lys
450 455 460
Ile Thr Cys Ser Ala His Arg Glu Pro Arg Leu Val Gly Gly Asp Ile
465 470 475 480
Pro Cys Ser Gly Arg Val Glu Val Lys His Gly Asp Thr Trp Gly Ser
485 490 495
Ile Cys Asp Ser Asp Phe Ser Leu Glu Ala Ala Ser Val Leu Cys Arg
500 505 510
Glu Leu Gln Cys Gly Thr Val Val Ser Ile Leu Gly Gly Ala His Phe
515 520 525
Gly Glu Gly Asn Gly Gln Ile Trp Ala Glu Glu Phe Gln Cys Glu Gly
530 535 540
His Glu Ser His Leu Ser Leu Cys Pro Val Ala Pro Arg Pro Glu Gly
545 550 555 560
Thr Cys Ser His Ser Arg Asp Val Gly Val Val Cys Ser Arg Tyr Thr
565 570 575
Glu Ile Arg Leu Val Asn Gly Lys Thr Pro Cys Glu Gly Arg Val Glu
580 585 590
Leu Lys Thr Leu Gly Ala Trp Gly Ser Leu Cys Asn Ser His Trp Asp
595 600 605
Ile Glu Asp Ala His Val Leu Cys Gln Gln Leu Lys Cys Gly Val Ala
610 615 620
Leu Ser Thr Pro Gly Gly Ala Arg Phe Gly Lys Gly Asn Gly Gln Ile
625 630 635 640
Trp Arg His Met Phe His Cys Thr Gly Thr Glu Gln His Met Gly Asp
645 650 655
Cys Pro Val Thr Ala Leu Gly Ala Ser Leu Cys Pro Ser Glu Gln Val
660 665 670
Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln Thr Leu Ser Ser Cys
675 680 685
Asn Ser Ser Ser Leu Gly Pro Thr Arg Pro Thr Ile Pro Glu Glu Ser
690 695 700
Ala Val Ala Cys Ile Glu Ser Gly Gln Leu Arg Leu Val Asn Gly Gly
705 710 715 720
Gly Arg Cys Ala Gly Arg Val Glu Ile Tyr His Glu Gly Ser Trp Gly
725 730 735
Thr Ile Cys Asp Asp Ser Trp Asp Leu Ser Asp Ala His Val Val Cys
740 745 750
Arg Gln Leu Gly Cys Gly Glu Ala Ile Asn Ala Thr Gly Ser Ala His
755 760 765
Phe Gly Glu Gly Thr Gly Pro Ile Trp Leu Asp Glu Met Lys Cys Asn
770 775 780
Gly Lys Glu Ser Arg Ile Trp Gln Cys His Ser His Gly Trp Gly Gln
785 790 795 800
Gln Asn Cys Arg His Lys Glu Asp Ala Gly Val Ile Cys Ser Glu Phe
805 810 815
Met Ser Leu Arg Leu Thr Ser Glu Ala Ser Arg Glu Ala Cys Ala Gly
820 825 830
Arg Leu Glu Val Phe Tyr Asn Gly Ala Trp Gly Thr Val Gly Lys Ser
835 840 845
Ser Met Ser Glu Thr Thr Val Gly Val Val Cys Arg Gln Leu Gly Cys
850 855 860
Ala Asp Lys Gly Lys Ile Asn Pro Ala Ser Leu Asp Lys Ala Met Ser
865 870 875 880
Ile Pro Met Trp Val Asp Asn Val Gln Cys Pro Lys Gly Pro Asp Thr
885 890 895
Leu Trp Gln Cys Pro Ser Ser Pro Trp Glu Lys Arg Leu Ala Ser Pro
900 905 910
Ser Glu Glu Thr Trp Ile Thr Cys Asp Asn Lys Ile Arg Leu Gln Glu
915 920 925
Gly Pro Thr Ser Cys Ser Gly Arg Val Glu Ile Trp His Gly Gly Ser
930 935 940
Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu Asp Asp Ala Gln Val
945 950 955 960
Val Cys Gln Gln Leu Gly Cys Gly Pro Ala Leu Lys Ala Phe Lys Glu
965 970 975
Ala Glu Phe Gly Gln Gly Thr Gly Pro Ile Trp Leu Asn Glu Val Lys
980 985 990
Cys Lys Gly Asn Glu Ser Ser Leu Trp Asp Cys Pro Ala Arg Arg Trp
995 1000 1005
Gly His Ser Glu Cys Gly His Lys Glu Asp Ala Ala Val Asn Cys
1010 1015 1020
Thr Asp Ile Ser Val Gln Lys Thr Pro Gln Lys Ala Thr Thr Gly
1025 1030 1035
Arg Ser Ser Arg Gln Ser Ser Phe Ile Ala Val Gly Ile Leu Gly
1040 1045 1050
Val Val Leu Leu Ala Ile Phe Val Ala Leu Phe Phe Leu Thr Lys
1055 1060 1065
Lys Arg Arg Gln Arg Gln Arg Leu Ala Val Ser Ser Arg Gly Glu
1070 1075 1080
Asn Leu Val His Gln Ile Gln Tyr Arg Glu Met Asn Ser Cys Leu
1085 1090 1095
Asn Ala Asp Asp Leu Asp Leu Met Asn Ser Ser Gly Leu Trp Val
1100 1105 1110
Leu Gly Gly Ser Ile Ala Gln Gly Phe Arg Ser Val Ala Ala Val
1115 1120 1125
Glu Ala Gln Thr Phe Tyr Phe Asp Lys Gln Leu Lys Lys Ser Lys
1130 1135 1140
Asn Val Ile Gly Ser Leu Asp Ala Tyr Asn Gly Gln Glu
1145 1150 1155
<210> SEQ ID NO 9
<211> LENGTH: 5193
<212> TYPE: DNA
<213> ORGANISM: sus scrofa
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/NM_214346
<309> DATABASE ENTRY DATE: 2004-08-05
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(5193)
<400> SEQUENCE: 9
atggacttcc tgctcctgct cctcctcctg gcttcatctg ctctagcagg cctggcctcg 60
tggacggttt ccagccccga gaccgtgcag ggcatcaagg gctcctgcct catcatcccc 120
tgcaccttcg gcttcccggc caacgtggag gtgccccatg gcatcacagc catctggtac 180
tatgactact caggcaagcg cctggtagtg agccactcca ggaacccaaa ggtggtggag 240
aaccacttcc aaggccgggc cctgctgttg gggcaggttg aacagaggac gtgcagcctg 300
ctgctgaagg acctgcagcc ccaggactcg ggctcctata acttccgctt tgagatcagc 360
gagggcaacc gctggtcaga tgtcaaaggc acagttgtca ccgtgacaga ggtgcccagc 420
gtgcccacca ttgccttgcc agccaagctg catgagggca tggaggtgga cttcaactgc 480
tccactccct atgtgtgccc gacggagccg gtcaacctac agtggcaagg ccaggatccc 540
acccgctccg tcacctccca cctccagaag cttgagccct cgggcaccag ccacatggag 600
accctgcaca tggccctgtc ctggcaggac catggccgga tcctgagctg ccaggtctca 660
gcagccgaac gcaggatgca gaaggagatt cacctccaag tgcagtatgc ccccaagggt 720
gtggagatcc ttttcagcca ctccggacgg aacgtccttc caggtgatct ggtcaccctc 780
agctgccagg tgaatagcag caaccctcag gtcagttccg tgcagtgggt caaggatggg 840
acgaagctca aagaccagaa acgtgtactg cagttgcgcc gggcagcctg ggctgatgct 900
ggcgtctaca cctgccaagc cgggaatgcc gtgggctctt cagtctcacc cccggtcagc 960
ctccacgtct tcatggctga ggtccaggta agccctgtgg gctccatcct ggagaaccag 1020
acggtgacgc tggcctgcaa tacacctaag gaagcgccca gcgagctgcg ctacagctgg 1080
tacaagaacc acgccctgct ggagggctct cacagccgca ccctccggct gcactcagtt 1140
accagggcgg attcgggctt ctacttctgc gaggtgcaga acgcccgggg cagagagcgc 1200
tctccccctg tcagcgtggt ggtcagccac ccacccctca ccccggacct aactgccttc 1260
ctggagacac aggcggggct ggtgggcatc ctccaatgct ctgtggtcag cgagccccca 1320
gctactctgg tgttgtcaca cgggggcctc atcttggcct ctacctccgg ggagggtgac 1380
cacagcccac gcttcagtgt cgcctctgcc cccaactccc tgcgcctgga gattcaagac 1440
ctggggccaa cagacagtgg ggaatacatg tgctcagcca gcagttctct tgggaatgcg 1500
tcctccaccc tggacttcca tgccaatgca gcccgcctcc tcatcagccc agcagcagag 1560
gtggtggaag ggcaggcggt gacactgagc tgcaggagca gcctgagcct gatgcctgac 1620
acccgttttt cctggtacct gaacggggcc ctgattctcg aggggcccag cagcagcctc 1680
ctgctcccag cagcctccag cacagatgcc ggctcatacc actgccgggc ccagaacagc 1740
cacagcacca gcgggccctc ctcacctgct gttctcaccg tgctctacgc cccacgccag 1800
cccgtgttca ctgcccagct ggaccctgat actgcaggag ctggggccgg acgccaaggc 1860
ctcctcttgt gccgtgtgga cagcgacccc ccagcccagc tgcagctgct ccacaggggc 1920
cgtgttgtgg cctcttctct gtcatggggg ggcggctgct gcacctgcgg aggctgtttc 1980
caccgcatga aggtcaccaa agcacccaac ctactgcgtg tagagatccg agacccggtg 2040
ctggaggatg agggtgtgta cctgtgcgag gccagcagcg ccctgggcaa cgcctccgcc 2100
tctgcaacct tggatgccca ggccactgtc ctggtcatca caccgtcaca cacgctgcag 2160
gaaggcattg aagccaacct gacttgcaac gtgagccgtg aagccagcgg ccctgccaac 2220
ttctcctggt tccgagatgg ggcgctatgg gcccagggcc ctctggacac cgtgacgctg 2280
ctacctgtgg ccagaactga tgctgccctc tatgcttgcc gcatcgtcac cgaggctggt 2340
gctggcctct ccacccctgt ggccctgaat gtgctctatc cccccgatcc tccaaagttg 2400
tcagccctcc tggacgtgga ccagggccac acggctgtgt tcgtctgtac tgtggacagt 2460
cgccctcttg cccagttggc cctgttccgt ggggaacacc tcctggccgc cagctcggca 2520
ctccggctcc cccctcgtgg ccgcctccag gccaaagcct cggccaactc cttgcagcta 2580
gaggtccgag acttgagcct tggggactct ggcagctacc actgtgaggc caccaacatc 2640
cttggatcag ccaacacttc tcttaccttc caggtccgag gagcctgggt ccgggtgtca 2700
ccgtcgcctg agctccagga gggccaggct gtggtcctga gctgccaggt acccataggg 2760
gtcctggagg ggacctcata tcgttggtat cgggatggcc agcccctcca ggagtccact 2820
tcggccacgc tccgttttgc agccataact ctgagccagg ctggagccta ccattgccaa 2880
gcccaagctc caggctcagc caccacggac ctggctgccc ctgtcagcct ccacgtgacc 2940
tacgcacctc gccaggccac actcaccacc ctgatggact caggcctcgg gcgactgggc 3000
ctccttctgt gccgtgtgaa cagtgaccct cctgcccagc tccgactgct ccatgggagc 3060
cgcctcgtgg cctctactct acaaggtgtg gaggagcttg caggcagctc tccccgccta 3120
caggtggcca cagcccccaa cacgctgcgc ctggagatcc acaacgcagt gctggaggat 3180
gaaggcgtct acacctgcga ggccaccaac accctgggtc agaccttggc ctccgccgcc 3240
ttcgatgccc aggctatgag agtgcaggtg tggcccaatg ccaccgtgca agaggggcag 3300
ctggtgaacc tgacctgcct tgtatggacc acgcacctgg cccagctcac ctacacgtgg 3360
taccgagacc agcagcagct cccaggtgct gcccactcca tcctcctgcc caatgtcact 3420
gtcacagatg ccgcctccta ccgctgtggc atattgatcc ctggccaggc actccgcctc 3480
tccagacctg tcgccctgga tgtcctctac gcaccccgca gactgcgcct gacccatctc 3540
ttggagagcc gtggtgggca gctggccgtg gtgctgtgca ctgtggacag tcgcccagct 3600
gcccagctga ccctcagcca tgctggccgc ctcctggcct cctcaaccgc agcctctgtc 3660
cccaacaccc tgcgcctgga gctgtgggag ccccggccca gtgatgaggg tctctacagc 3720
tgctcggccc gcagtcctct gggccaggcc aacacatccc tggagctgcg gctagagggc 3780
gtgcaggtgg cactggctcc atcggccact gtgccggagg gggcccctgt cacagtgacc 3840
tgtgaagacc ctgctgcccg cccacccact ctctatgtct ggtaccacaa cagccgttgg 3900
ctgcaggagg ggtcggctgc ctccctctcg tttccagcgg ctacacgggc tcacgcgggc 3960
gcctatacct gccaggtcca ggatgcccag ggcacacgca tctcccagcc cgcagcactg 4020
cacatcctct atgcccctcg ggatgctgtc ctttcctcct tctgggactc aagggccagc 4080
cctatggccg tggtacagtg cactgtggac agcgagccac ctgccgagat gaccctgtcc 4140
catgatggca aggtgctggc caccagccat ggggtccacg gcttagcagt ggggacaggc 4200
catgtccagg tggcccgcaa cgccctgcag ctgcgggtgc agaatgtgcc ctcacgtgac 4260
aaggacacct acgtctgcat ggaccgcaac tccttgggct cagtcagcac catggggcag 4320
ctgcagccag aaggtgtgca cgtggtagct gagccagggc tggatgtgcc tgaaggcaca 4380
gcgctgaacc tgagctgtcg cctccctagt ggccctgggc acataggcaa ctccaccttt 4440
gcttggttcc ggaacggtcg gcagctacac acagagtctg tgcccaccct taccttcacc 4500
catgtggccc gcgcccaagc tggcttgtac cactgccagg ctgagctccc cgccggggct 4560
gccacctctg ctccagtctt gctccgggtg ctctaccctc ccaagacgcc caccatgact 4620
gtttttgtgg agcccgaggg tggcatccag ggcattctgg actgccgagt ggacagtgag 4680
cccctagcca gcctgaccct ccacctgggc agtcggctgg tggcctccag ccagcctcag 4740
gctgcccctg ccaagccgca catccgcgtc tcagccagtc ccaatgcctt gcgagtggac 4800
atggaggagc tgaagcccag tgaccagggg gagtatgtgt gctcggcctc caatgccctg 4860
ggctctgcct ctgctgccac ctacttcgga accagagccc tgcatcgcct gcatctgttc 4920
cagcaccttc tctggttcct ggggctgctg gcgagcctcc tcttcctact gttgggcctg 4980
ggggtctggt acgcctggag acggggaaat ttttacaagc tgagaatggg cgaatattca 5040
gtagagatgg tatctcggaa ggaaaccacg cagatgtcca ctgaccagga agaagttact 5100
ggaatcggtg atgatgcggg ctctgtgaac caggcggcat ttgatcctgc ccacctctgt 5160
gaaaacacac agtctgtgaa aagcacagtc tga 5193
<210> SEQ ID NO 10
<211> LENGTH: 1730
<212> TYPE: PRT
<213> ORGANISM: sus scrofa
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/NP_999511
<309> DATABASE ENTRY DATE: 2004-08-05
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1730)
<400> SEQUENCE: 10
Met Asp Phe Leu Leu Leu Leu Leu Leu Leu Ala Ser Ser Ala Leu Ala
1 5 10 15
Gly Leu Ala Ser Trp Thr Val Ser Ser Pro Glu Thr Val Gln Gly Ile
20 25 30
Lys Gly Ser Cys Leu Ile Ile Pro Cys Thr Phe Gly Phe Pro Ala Asn
35 40 45
Val Glu Val Pro His Gly Ile Thr Ala Ile Trp Tyr Tyr Asp Tyr Ser
50 55 60
Gly Lys Arg Leu Val Val Ser His Ser Arg Asn Pro Lys Val Val Glu
65 70 75 80
Asn His Phe Gln Gly Arg Ala Leu Leu Leu Gly Gln Val Glu Gln Arg
85 90 95
Thr Cys Ser Leu Leu Leu Lys Asp Leu Gln Pro Gln Asp Ser Gly Ser
100 105 110
Tyr Asn Phe Arg Phe Glu Ile Ser Glu Gly Asn Arg Trp Ser Asp Val
115 120 125
Lys Gly Thr Val Val Thr Val Thr Glu Val Pro Ser Val Pro Thr Ile
130 135 140
Ala Leu Pro Ala Lys Leu His Glu Gly Met Glu Val Asp Phe Asn Cys
145 150 155 160
Ser Thr Pro Tyr Val Cys Pro Thr Glu Pro Val Asn Leu Gln Trp Gln
165 170 175
Gly Gln Asp Pro Thr Arg Ser Val Thr Ser His Leu Gln Lys Leu Glu
180 185 190
Pro Ser Gly Thr Ser His Met Glu Thr Leu His Met Ala Leu Ser Trp
195 200 205
Gln Asp His Gly Arg Ile Leu Ser Cys Gln Val Ser Ala Ala Glu Arg
210 215 220
Arg Met Gln Lys Glu Ile His Leu Gln Val Gln Tyr Ala Pro Lys Gly
225 230 235 240
Val Glu Ile Leu Phe Ser His Ser Gly Arg Asn Val Leu Pro Gly Asp
245 250 255
Leu Val Thr Leu Ser Cys Gln Val Asn Ser Ser Asn Pro Gln Val Ser
260 265 270
Ser Val Gln Trp Val Lys Asp Gly Thr Lys Leu Lys Asp Gln Lys Arg
275 280 285
Val Leu Gln Leu Arg Arg Ala Ala Trp Ala Asp Ala Gly Val Tyr Thr
290 295 300
Cys Gln Ala Gly Asn Ala Val Gly Ser Ser Val Ser Pro Pro Val Ser
305 310 315 320
Leu His Val Phe Met Ala Glu Val Gln Val Ser Pro Val Gly Ser Ile
325 330 335
Leu Glu Asn Gln Thr Val Thr Leu Ala Cys Asn Thr Pro Lys Glu Ala
340 345 350
Pro Ser Glu Leu Arg Tyr Ser Trp Tyr Lys Asn His Ala Leu Leu Glu
355 360 365
Gly Ser His Ser Arg Thr Leu Arg Leu His Ser Val Thr Arg Ala Asp
370 375 380
Ser Gly Phe Tyr Phe Cys Glu Val Gln Asn Ala Arg Gly Arg Glu Arg
385 390 395 400
Ser Pro Pro Val Ser Val Val Val Ser His Pro Pro Leu Thr Pro Asp
405 410 415
Leu Thr Ala Phe Leu Glu Thr Gln Ala Gly Leu Val Gly Ile Leu Gln
420 425 430
Cys Ser Val Val Ser Glu Pro Pro Ala Thr Leu Val Leu Ser His Gly
435 440 445
Gly Leu Ile Leu Ala Ser Thr Ser Gly Glu Gly Asp His Ser Pro Arg
450 455 460
Phe Ser Val Ala Ser Ala Pro Asn Ser Leu Arg Leu Glu Ile Gln Asp
465 470 475 480
Leu Gly Pro Thr Asp Ser Gly Glu Tyr Met Cys Ser Ala Ser Ser Ser
485 490 495
Leu Gly Asn Ala Ser Ser Thr Leu Asp Phe His Ala Asn Ala Ala Arg
500 505 510
Leu Leu Ile Ser Pro Ala Ala Glu Val Val Glu Gly Gln Ala Val Thr
515 520 525
Leu Ser Cys Arg Ser Ser Leu Ser Leu Met Pro Asp Thr Arg Phe Ser
530 535 540
Trp Tyr Leu Asn Gly Ala Leu Ile Leu Glu Gly Pro Ser Ser Ser Leu
545 550 555 560
Leu Leu Pro Ala Ala Ser Ser Thr Asp Ala Gly Ser Tyr His Cys Arg
565 570 575
Ala Gln Asn Ser His Ser Thr Ser Gly Pro Ser Ser Pro Ala Val Leu
580 585 590
Thr Val Leu Tyr Ala Pro Arg Gln Pro Val Phe Thr Ala Gln Leu Asp
595 600 605
Pro Asp Thr Ala Gly Ala Gly Ala Gly Arg Gln Gly Leu Leu Leu Cys
610 615 620
Arg Val Asp Ser Asp Pro Pro Ala Gln Leu Gln Leu Leu His Arg Gly
625 630 635 640
Arg Val Val Ala Ser Ser Leu Ser Trp Gly Gly Gly Cys Cys Thr Cys
645 650 655
Gly Gly Cys Phe His Arg Met Lys Val Thr Lys Ala Pro Asn Leu Leu
660 665 670
Arg Val Glu Ile Arg Asp Pro Val Leu Glu Asp Glu Gly Val Tyr Leu
675 680 685
Cys Glu Ala Ser Ser Ala Leu Gly Asn Ala Ser Ala Ser Ala Thr Leu
690 695 700
Asp Ala Gln Ala Thr Val Leu Val Ile Thr Pro Ser His Thr Leu Gln
705 710 715 720
Glu Gly Ile Glu Ala Asn Leu Thr Cys Asn Val Ser Arg Glu Ala Ser
725 730 735
Gly Pro Ala Asn Phe Ser Trp Phe Arg Asp Gly Ala Leu Trp Ala Gln
740 745 750
Gly Pro Leu Asp Thr Val Thr Leu Leu Pro Val Ala Arg Thr Asp Ala
755 760 765
Ala Leu Tyr Ala Cys Arg Ile Val Thr Glu Ala Gly Ala Gly Leu Ser
770 775 780
Thr Pro Val Ala Leu Asn Val Leu Tyr Pro Pro Asp Pro Pro Lys Leu
785 790 795 800
Ser Ala Leu Leu Asp Val Asp Gln Gly His Thr Ala Val Phe Val Cys
805 810 815
Thr Val Asp Ser Arg Pro Leu Ala Gln Leu Ala Leu Phe Arg Gly Glu
820 825 830
His Leu Leu Ala Ala Ser Ser Ala Leu Arg Leu Pro Pro Arg Gly Arg
835 840 845
Leu Gln Ala Lys Ala Ser Ala Asn Ser Leu Gln Leu Glu Val Arg Asp
850 855 860
Leu Ser Leu Gly Asp Ser Gly Ser Tyr His Cys Glu Ala Thr Asn Ile
865 870 875 880
Leu Gly Ser Ala Asn Thr Ser Leu Thr Phe Gln Val Arg Gly Ala Trp
885 890 895
Val Arg Val Ser Pro Ser Pro Glu Leu Gln Glu Gly Gln Ala Val Val
900 905 910
Leu Ser Cys Gln Val Pro Ile Gly Val Leu Glu Gly Thr Ser Tyr Arg
915 920 925
Trp Tyr Arg Asp Gly Gln Pro Leu Gln Glu Ser Thr Ser Ala Thr Leu
930 935 940
Arg Phe Ala Ala Ile Thr Leu Ser Gln Ala Gly Ala Tyr His Cys Gln
945 950 955 960
Ala Gln Ala Pro Gly Ser Ala Thr Thr Asp Leu Ala Ala Pro Val Ser
965 970 975
Leu His Val Thr Tyr Ala Pro Arg Gln Ala Thr Leu Thr Thr Leu Met
980 985 990
Asp Ser Gly Leu Gly Arg Leu Gly Leu Leu Leu Cys Arg Val Asn Ser
995 1000 1005
Asp Pro Pro Ala Gln Leu Arg Leu Leu His Gly Ser Arg Leu Val
1010 1015 1020
Ala Ser Thr Leu Gln Gly Val Glu Glu Leu Ala Gly Ser Ser Pro
1025 1030 1035
Arg Leu Gln Val Ala Thr Ala Pro Asn Thr Leu Arg Leu Glu Ile
1040 1045 1050
His Asn Ala Val Leu Glu Asp Glu Gly Val Tyr Thr Cys Glu Ala
1055 1060 1065
Thr Asn Thr Leu Gly Gln Thr Leu Ala Ser Ala Ala Phe Asp Ala
1070 1075 1080
Gln Ala Met Arg Val Gln Val Trp Pro Asn Ala Thr Val Gln Glu
1085 1090 1095
Gly Gln Leu Val Asn Leu Thr Cys Leu Val Trp Thr Thr His Leu
1100 1105 1110
Ala Gln Leu Thr Tyr Thr Trp Tyr Arg Asp Gln Gln Gln Leu Pro
1115 1120 1125
Gly Ala Ala His Ser Ile Leu Leu Pro Asn Val Thr Val Thr Asp
1130 1135 1140
Ala Ala Ser Tyr Arg Cys Gly Ile Leu Ile Pro Gly Gln Ala Leu
1145 1150 1155
Arg Leu Ser Arg Pro Val Ala Leu Asp Val Leu Tyr Ala Pro Arg
1160 1165 1170
Arg Leu Arg Leu Thr His Leu Leu Glu Ser Arg Gly Gly Gln Leu
1175 1180 1185
Ala Val Val Leu Cys Thr Val Asp Ser Arg Pro Ala Ala Gln Leu
1190 1195 1200
Thr Leu Ser His Ala Gly Arg Leu Leu Ala Ser Ser Thr Ala Ala
1205 1210 1215
Ser Val Pro Asn Thr Leu Arg Leu Glu Leu Trp Glu Pro Arg Pro
1220 1225 1230
Ser Asp Glu Gly Leu Tyr Ser Cys Ser Ala Arg Ser Pro Leu Gly
1235 1240 1245
Gln Ala Asn Thr Ser Leu Glu Leu Arg Leu Glu Gly Val Gln Val
1250 1255 1260
Ala Leu Ala Pro Ser Ala Thr Val Pro Glu Gly Ala Pro Val Thr
1265 1270 1275
Val Thr Cys Glu Asp Pro Ala Ala Arg Pro Pro Thr Leu Tyr Val
1280 1285 1290
Trp Tyr His Asn Ser Arg Trp Leu Gln Glu Gly Ser Ala Ala Ser
1295 1300 1305
Leu Ser Phe Pro Ala Ala Thr Arg Ala His Ala Gly Ala Tyr Thr
1310 1315 1320
Cys Gln Val Gln Asp Ala Gln Gly Thr Arg Ile Ser Gln Pro Ala
1325 1330 1335
Ala Leu His Ile Leu Tyr Ala Pro Arg Asp Ala Val Leu Ser Ser
1340 1345 1350
Phe Trp Asp Ser Arg Ala Ser Pro Met Ala Val Val Gln Cys Thr
1355 1360 1365
Val Asp Ser Glu Pro Pro Ala Glu Met Thr Leu Ser His Asp Gly
1370 1375 1380
Lys Val Leu Ala Thr Ser His Gly Val His Gly Leu Ala Val Gly
1385 1390 1395
Thr Gly His Val Gln Val Ala Arg Asn Ala Leu Gln Leu Arg Val
1400 1405 1410
Gln Asn Val Pro Ser Arg Asp Lys Asp Thr Tyr Val Cys Met Asp
1415 1420 1425
Arg Asn Ser Leu Gly Ser Val Ser Thr Met Gly Gln Leu Gln Pro
1430 1435 1440
Glu Gly Val His Val Val Ala Glu Pro Gly Leu Asp Val Pro Glu
1445 1450 1455
Gly Thr Ala Leu Asn Leu Ser Cys Arg Leu Pro Ser Gly Pro Gly
1460 1465 1470
His Ile Gly Asn Ser Thr Phe Ala Trp Phe Arg Asn Gly Arg Gln
1475 1480 1485
Leu His Thr Glu Ser Val Pro Thr Leu Thr Phe Thr His Val Ala
1490 1495 1500
Arg Ala Gln Ala Gly Leu Tyr His Cys Gln Ala Glu Leu Pro Ala
1505 1510 1515
Gly Ala Ala Thr Ser Ala Pro Val Leu Leu Arg Val Leu Tyr Pro
1520 1525 1530
Pro Lys Thr Pro Thr Met Thr Val Phe Val Glu Pro Glu Gly Gly
1535 1540 1545
Ile Gln Gly Ile Leu Asp Cys Arg Val Asp Ser Glu Pro Leu Ala
1550 1555 1560
Ser Leu Thr Leu His Leu Gly Ser Arg Leu Val Ala Ser Ser Gln
1565 1570 1575
Pro Gln Ala Ala Pro Ala Lys Pro His Ile Arg Val Ser Ala Ser
1580 1585 1590
Pro Asn Ala Leu Arg Val Asp Met Glu Glu Leu Lys Pro Ser Asp
1595 1600 1605
Gln Gly Glu Tyr Val Cys Ser Ala Ser Asn Ala Leu Gly Ser Ala
1610 1615 1620
Ser Ala Ala Thr Tyr Phe Gly Thr Arg Ala Leu His Arg Leu His
1625 1630 1635
Leu Phe Gln His Leu Leu Trp Phe Leu Gly Leu Leu Ala Ser Leu
1640 1645 1650
Leu Phe Leu Leu Leu Gly Leu Gly Val Trp Tyr Ala Trp Arg Arg
1655 1660 1665
Gly Asn Phe Tyr Lys Leu Arg Met Gly Glu Tyr Ser Val Glu Met
1670 1675 1680
Val Ser Arg Lys Glu Thr Thr Gln Met Ser Thr Asp Gln Glu Glu
1685 1690 1695
Val Thr Gly Ile Gly Asp Asp Ala Gly Ser Val Asn Gln Ala Ala
1700 1705 1710
Phe Asp Pro Ala His Leu Cys Glu Asn Thr Gln Ser Val Lys Ser
1715 1720 1725
Thr Val
1730
<210> SEQ ID NO 11
<211> LENGTH: 6387
<212> TYPE: DNA
<213> ORGANISM: mus musculus
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/NM_011426
<309> DATABASE ENTRY DATE: 2000-01-25
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(6387)
<400> SEQUENCE: 11
agacaagatt aggcctagag taagtctatg aaacacagag aaaggggaca gcataggggt 60
taagaaatga ggtctttcaa aatctcaggg ggcaatgagg agttttttga gagaggaagg 120
actctttaaa ggaagttgaa ggaggattct gtgaacttga gaccaccctg agctgccaag 180
ttgagaactt tgtctacaaa caagccaggc agcctcagcg tgtgctcagt ccgacttgta 240
gctggagagg caggagacca atttccggtg cttacggtgc ttgctggatg ccctggagta 300
agtgacaggg tctcactgga ctccaggttc tgttggtttg agtaatagga ggcggcaggg 360
gagaagtgaa gagagacatg cactgctgat ctgccttgag gctgtgtcct taaggggtgg 420
agccaagggg cacagaagac tctctgggac atgccaccaa gtgagagcat ttccaatcac 480
tccctgagcc aggaacaggg gcttctggtt ccctgctggt ggctgccaca gcagtccttc 540
ctgttgggtt gaccaacaca gcaggtgaga taaaccctat agacttgggc cctggagtgc 600
tccaggcagt ctctgtgtgc ctacccaccc ggcttcccta ggcacctgaa tgcacctggg 660
cactgggatg tgtgtcctgt tctccctgct cctgctggcc tctgtcttct cactaggcca 720
gaccacatgg ggtgtctcca gtcccaagaa tgtgcagggc ttgtcgggat cctgcctgct 780
cattccctgc atcttcagct accctgccga tgtcccagtg tccaatggca tcacagccat 840
ctggtactat gactactcgg gcaagcggca ggtggtaatc cactcagggg accccaagct 900
ggtggacaag cgtttcaggg gtcgagctga actgatgggg aacatggacc acaaggtgtg 960
caacctgttg ctcaaagact tgaagcctga agactctggc acctacaact tccgctttga 1020
gatcagtgat agcaaccgct ggttagatgt caaaggcacc acggtcactg tgacaacgga 1080
tcccagcccc cccactatta ccattcctga ggagctgcgt gaaggcatgg agaggaactt 1140
caactgttcc acaccctacc tgtgcctgca ggagaagcaa gtcagcctgc agtggcgagg 1200
ccaggacccc acccactctg tcacctccag cttccagagc ctcgagccca ctggcgtcta 1260
tcaccagacg accctacata tggccctatc ctggcaggac cacggtcgga ccctgctctg 1320
ccagttctca ttgggcgcac acagtagtcg gaaagaggtt tacctgcaag tgccacatgc 1380
ccccaaaggt gtggagatcc tcctcagctc ctcagggagg aacatccttc ccggggatcc 1440
agtcacactc acctgcagag tgaacagcag ctatcctgct gtcagtgccg tgcagtgggc 1500
cagggacgga gtgaacctcg gagtcacggg acatgtgctt cggctgttct cagcagcctg 1560
gaatgattct ggggcctaca cctgccaagc aacaaatgat atgggctctc tggtgtcatc 1620
cccgctcagc ctccatgttt ttatggctga agtcaaaatg aaccccgcag ggcccgtctt 1680
ggaaaatgag acagtgactc tgctctgtag cacgccgaag gaggctcccc aggagctccg 1740
ctatagctgg tacaagaacc acattctcct ggaagatgcc catgcctcaa ccttgcacct 1800
gcctgcagtc accagggctg atactggctt ctacttctgt gaagtgcaga atgcccaggg 1860
cagtgagcgc tccagtccat tgagtgtggt ggtcagatat ccacccctta ctccagacct 1920
gaccaccttc ctggagacac aggccggact tgtgggcatc ttgcattgct ccgtggtcag 1980
tgagcccctg gctactgtgg tgctgtcaca cggaggcctc acgttggcct ccaactctgg 2040
agaaaatgac ttcaaccccc gattcaggat ctcctctgcc cccaactccc tgcgcctaga 2100
aatccgagac ttgcagccag cagacagcgg agagtacaca tgcttagctg tcaactccct 2160
tggaaactca acgtccagcc tagacttcta tgctaatgtg gcccgactcc tcatcaaccc 2220
ttcagcagag gttgtggaag ggcaggcggt gaccctgagc tgcaggagtg gcctgagccc 2280
agctcctgac actcgcttct cctggtacct gaacggagct ctacttctgg aaggatccag 2340
cagcagcctc ctgcttcctg cggcttccag cactgatgcg ggctcatact actgtaggac 2400
gcaggctggc cccaacacca gcggcccctc cctgcctact gtcctcactg tgttctatcc 2460
cccaagaaag cccacattca ctgccaggct ggatttggat acctctggag tcggggatgg 2520
acgacggggc atcctcttgt gccacgtaga cagcgatccc ccagcccagc tacggcttct 2580
ccacaaaggc catgttgtgg ccacttctct gccatcaagg tgtgggagct gttcccagcg 2640
cacaaaagtc agcagaacct ccaactcact gcacgtggag atccagaagc ctgtattaga 2700
ggatgagggc gtgtacctgt gtgaggctag caacacattg ggcaactcct cagccgcagc 2760
ctctttcaat gctaaggcca ctgtactggt catcacaccg tcaaatacac tgcgtgaagg 2820
cacagaggcc aacctaactt gcaacgtgaa ccaggaggtt gctgtcagcc ctgccaactt 2880
ctcctggttc cggaatggag tgctgtggac ccagggatca ctggagactg tgaggctgca 2940
gcctgtggcc agaactgatg ctgctgtcta tgcctgccgc ctcctcaccg aggatggggc 3000
tcagctctcg gctcctgtgg tcctaagtgt gctgtatgcc ccagaccctc caaagctgtc 3060
agccctccta gatgtgggtc agggccacat ggccgtgttc atctgcactg tggacagcta 3120
tcccctggct cacctgtctc tgttccgtgg ggaccatctc ctggccacca acttggaacc 3180
ccagcgtccc tcccatggca ggatccaggc caaggccaca gccaactccc tgcagctaga 3240
ggtccgagaa ctaggtcttg tggactctgg aaactaccac tgtgaagcca ccaatattct 3300
tgggtcagcc aacagttcac tcttcttcca ggtcagagga gcctgggtcc aggtttcacc 3360
atcacctgag ctccgggagg gccaggctgt ggtcctgagc tgccaggtgc ccacaggagt 3420
ctctgagggg acctcataca gctggtatca ggatggccgc cccctccagg agtcaacctc 3480
atctacactc cgcattgcag ccataagtct gaggcaagct ggtgcctacc attgccaagc 3540
tcaggcccca gacacagcta ttgccagcct ggctgcccct gtcagcctcc atgtgtccta 3600
taccccacgt catgttacac tcagtgccct gctgagcacg gaccctgagc gactaggcca 3660
cctggtgtgc agtgtacaaa gtgaccctcc agcgcagctg caactgtttc accggaatcg 3720
cctcgtggcc tctaccctac aaggcgcgga cgaattggca ggcagtaatc cccggctgca 3780
tgtgactgtg ctccccaatg agctgcgcct gcagatccac tttccagagc tggaggatga 3840
cgggacctat acatgcgaag ccagcaacac actgggccag gcctcggctg cagctgactt 3900
cgatgcccag gctgtgcgag tgactgtgtg gcccaatgcc actgtgcaag aggggcagca 3960
ggtgaacctg acctgcttgg tgtggagcac ccaccaggac tcactcagct acacatggta 4020
caagggcggg caacaactcc ttggtgccag atccatcacc ctgcccagtg ttaaggtttt 4080
ggatgctacc tcctaccgct gtggtgtggg gctccccggc cacgcacccc atctctccag 4140
acccgtgacc ctggatgtcc tccatgctcc ccgaaacctg cggctgacct acctcctaga 4200
gacccagggc aggcagctgg ccctggtact gtgtacggtg gatagtcgtc cacctgccca 4260
gctaactctc agccatggtg accagcttgt agcctcctca actgaagcct ctgtccccaa 4320
caccctgcgc ctagagcttc aggatccaag gcctagtaat gaggggctct atagctgctc 4380
tgcccacagc ccattgggca aggccaacac gtccctggaa cttctgctgg aaggtgtccg 4440
agtgaaaatg aatccctctg gtagtgtacc cgagggagag cctgtcacag tgacctgcga 4500
ggaccctgct gccctctcat ccgccctcta tgcctggttt cacaatggcc attggcttca 4560
ggagggaccg gcttcctcac tccagttcct ggtgactaca cgggctcacg ctggtgctta 4620
cttttgccag gtgcatgata cacaaggcac acggagctcc agacctgcca gcctgcaaat 4680
tctctatgcc ccccgggatg ctgtcctgtc ttcctttcga gactcaagga ccaggctcat 4740
ggtcgtgatt cagtgcaccg tggacagtga gccacctgct gagatggtcc tatcccacaa 4800
tggcaaggtg ctagctgcca gccacgagcg tcacagctca gcatcaggga taggccacat 4860
ccaggtagcc cgaaatgctc ttcgactaca agtgcaagat gtgactctgg gtgatggcaa 4920
cacctatgtt tgcacagccc agaatacact gggctccatc agtaccaccc agaggcttct 4980
gacggagact gatatacgtg tgacagctga gccaggcttg gatgtgccag agggcacagc 5040
tctgaactta agctgcctcc tccctggtgg ctctgggccc acgggcaact cttccttcac 5100
gtggttctgg aatcgccacc gactacattc agctcctgtg cccacactct ccttcacccc 5160
tgtggtccgg gctcaggctg ggctgtacca ctgcagggct gatctcccca ccggggccac 5220
tacctctgct ccagttatgc tccgtgtcct ctatcccccc aagacgccca ctctcatagt 5280
gtttgtggag cctcagggtg gccaccaggg catcctcgac tgtcgagtgg acagtgagcc 5340
cctggccatc ctcactcttc accggggcag tcaactagta gcctccaacc aacttcacga 5400
tgctcccacc aagccccaca tccgagtcac tgctcctccc aatgccttga gagtggacat 5460
agaggagctc ggccctagca atcaagggga gtatgtgtgc actgcctcca acactctggg 5520
ctctgcctca gcctctgcct actttgggac cagagctctg caccaactgc agctgttcca 5580
gaggctgctc tgggtcctgg gatttctggc aggcttcctg tgcctgctgc tgggtctggt 5640
ggcctatcac acctggagaa agaagagttc taccaagctg aatgaggatg agaattcagc 5700
agagatggcc actaagaaaa atactatcca ggaggaagtg gttgctgctc tctgacaact 5760
caggtgctgt gaacaagatc ctgcctacct ctgtataagc agtacagaga catctggctt 5820
tcctgacctg cccgacttgc cttccaagcc tcttgatcct aagaaaaatg gacgaaggga 5880
ggtttggggt tggaggtcaa cctgccgcct ccagggctct gagacggact cagccatgtt 5940
gcccacgtct ctctgtgtgg ttttcctctg tatccctttg cctttctctt caaagctcac 6000
cttggacttt cttggtgggt tagagcaaca tccagtttct cacagacttt ctaagacggt 6060
ctgtaccagc caggatatca gtcaggttgc tctaacagag actcaataca gtgaccacag 6120
catgacaggg tcttagtttt ccctcctggc ctggttatgt tgttgtggta tcagaatcct 6180
tcttgcttga ttttctccat tccccaagtg ttgcctttga ttatgaagct caggtaactg 6240
cagtgcccat ggaccctaca gggagaagga agagtgaagg gaagacatac ccatccccat 6300
ggtccatgga ctgtgtgtgc aattgcaccc cacccaactt ctcatccgct agaaactggt 6360
cacataaaca taccatgctg aaaggga 6387
<210> SEQ ID NO 12
<211> LENGTH: 1695
<212> TYPE: PRT
<213> ORGANISM: mus musculus
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept
<309> DATABASE ENTRY DATE: 2000-01-25
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1695)
<400> SEQUENCE: 12
Met Cys Val Leu Phe Ser Leu Leu Leu Leu Ala Ser Val Phe Ser Leu
1 5 10 15
Gly Gln Thr Thr Trp Gly Val Ser Ser Pro Lys Asn Val Gln Gly Leu
20 25 30
Ser Gly Ser Cys Leu Leu Ile Pro Cys Ile Phe Ser Tyr Pro Ala Asp
35 40 45
Val Pro Val Ser Asn Gly Ile Thr Ala Ile Trp Tyr Tyr Asp Tyr Ser
50 55 60
Gly Lys Arg Gln Val Val Ile His Ser Gly Asp Pro Lys Leu Val Asp
65 70 75 80
Lys Arg Phe Arg Gly Arg Ala Glu Leu Met Gly Asn Met Asp His Lys
85 90 95
Val Cys Asn Leu Leu Leu Lys Asp Leu Lys Pro Glu Asp Ser Gly Thr
100 105 110
Tyr Asn Phe Arg Phe Glu Ile Ser Asp Ser Asn Arg Trp Leu Asp Val
115 120 125
Lys Gly Thr Thr Val Thr Val Thr Thr Asp Pro Ser Pro Pro Thr Ile
130 135 140
Thr Ile Pro Glu Glu Leu Arg Glu Gly Met Glu Arg Asn Phe Asn Cys
145 150 155 160
Ser Thr Pro Tyr Leu Cys Leu Gln Glu Lys Gln Val Ser Leu Gln Trp
165 170 175
Arg Gly Gln Asp Pro Thr His Ser Val Thr Ser Ser Phe Gln Ser Leu
180 185 190
Glu Pro Thr Gly Val Tyr His Gln Thr Thr Leu His Met Ala Leu Ser
195 200 205
Trp Gln Asp His Gly Arg Thr Leu Leu Cys Gln Phe Ser Leu Gly Ala
210 215 220
His Ser Ser Arg Lys Glu Val Tyr Leu Gln Val Pro His Ala Pro Lys
225 230 235 240
Gly Val Glu Ile Leu Leu Ser Ser Ser Gly Arg Asn Ile Leu Pro Gly
245 250 255
Asp Pro Val Thr Leu Thr Cys Arg Val Asn Ser Ser Tyr Pro Ala Val
260 265 270
Ser Ala Val Gln Trp Ala Arg Asp Gly Val Asn Leu Gly Val Thr Gly
275 280 285
His Val Leu Arg Leu Phe Ser Ala Ala Trp Asn Asp Ser Gly Ala Tyr
290 295 300
Thr Cys Gln Ala Thr Asn Asp Met Gly Ser Leu Val Ser Ser Pro Leu
305 310 315 320
Ser Leu His Val Phe Met Ala Glu Val Lys Met Asn Pro Ala Gly Pro
325 330 335
Val Leu Glu Asn Glu Thr Val Thr Leu Leu Cys Ser Thr Pro Lys Glu
340 345 350
Ala Pro Gln Glu Leu Arg Tyr Ser Trp Tyr Lys Asn His Ile Leu Leu
355 360 365
Glu Asp Ala His Ala Ser Thr Leu His Leu Pro Ala Val Thr Arg Ala
370 375 380
Asp Thr Gly Phe Tyr Phe Cys Glu Val Gln Asn Ala Gln Gly Ser Glu
385 390 395 400
Arg Ser Ser Pro Leu Ser Val Val Val Arg Tyr Pro Pro Leu Thr Pro
405 410 415
Asp Leu Thr Thr Phe Leu Glu Thr Gln Ala Gly Leu Val Gly Ile Leu
420 425 430
His Cys Ser Val Val Ser Glu Pro Leu Ala Thr Val Val Leu Ser His
435 440 445
Gly Gly Leu Thr Leu Ala Ser Asn Ser Gly Glu Asn Asp Phe Asn Pro
450 455 460
Arg Phe Arg Ile Ser Ser Ala Pro Asn Ser Leu Arg Leu Glu Ile Arg
465 470 475 480
Asp Leu Gln Pro Ala Asp Ser Gly Glu Tyr Thr Cys Leu Ala Val Asn
485 490 495
Ser Leu Gly Asn Ser Thr Ser Ser Leu Asp Phe Tyr Ala Asn Val Ala
500 505 510
Arg Leu Leu Ile Asn Pro Ser Ala Glu Val Val Glu Gly Gln Ala Val
515 520 525
Thr Leu Ser Cys Arg Ser Gly Leu Ser Pro Ala Pro Asp Thr Arg Phe
530 535 540
Ser Trp Tyr Leu Asn Gly Ala Leu Leu Leu Glu Gly Ser Ser Ser Ser
545 550 555 560
Leu Leu Leu Pro Ala Ala Ser Ser Thr Asp Ala Gly Ser Tyr Tyr Cys
565 570 575
Arg Thr Gln Ala Gly Pro Asn Thr Ser Gly Pro Ser Leu Pro Thr Val
580 585 590
Leu Thr Val Phe Tyr Pro Pro Arg Lys Pro Thr Phe Thr Ala Arg Leu
595 600 605
Asp Leu Asp Thr Ser Gly Val Gly Asp Gly Arg Arg Gly Ile Leu Leu
610 615 620
Cys His Val Asp Ser Asp Pro Pro Ala Gln Leu Arg Leu Leu His Lys
625 630 635 640
Gly His Val Val Ala Thr Ser Leu Pro Ser Arg Cys Gly Ser Cys Ser
645 650 655
Gln Arg Thr Lys Val Ser Arg Thr Ser Asn Ser Leu His Val Glu Ile
660 665 670
Gln Lys Pro Val Leu Glu Asp Glu Gly Val Tyr Leu Cys Glu Ala Ser
675 680 685
Asn Thr Leu Gly Asn Ser Ser Ala Ala Ala Ser Phe Asn Ala Lys Ala
690 695 700
Thr Val Leu Val Ile Thr Pro Ser Asn Thr Leu Arg Glu Gly Thr Glu
705 710 715 720
Ala Asn Leu Thr Cys Asn Val Asn Gln Glu Val Ala Val Ser Pro Ala
725 730 735
Asn Phe Ser Trp Phe Arg Asn Gly Val Leu Trp Thr Gln Gly Ser Leu
740 745 750
Glu Thr Val Arg Leu Gln Pro Val Ala Arg Thr Asp Ala Ala Val Tyr
755 760 765
Ala Cys Arg Leu Leu Thr Glu Asp Gly Ala Gln Leu Ser Ala Pro Val
770 775 780
Val Leu Ser Val Leu Tyr Ala Pro Asp Pro Pro Lys Leu Ser Ala Leu
785 790 795 800
Leu Asp Val Gly Gln Gly His Met Ala Val Phe Ile Cys Thr Val Asp
805 810 815
Ser Tyr Pro Leu Ala His Leu Ser Leu Phe Arg Gly Asp His Leu Leu
820 825 830
Ala Thr Asn Leu Glu Pro Gln Arg Pro Ser His Gly Arg Ile Gln Ala
835 840 845
Lys Ala Thr Ala Asn Ser Leu Gln Leu Glu Val Arg Glu Leu Gly Leu
850 855 860
Val Asp Ser Gly Asn Tyr His Cys Glu Ala Thr Asn Ile Leu Gly Ser
865 870 875 880
Ala Asn Ser Ser Leu Phe Phe Gln Val Arg Gly Ala Trp Val Gln Val
885 890 895
Ser Pro Ser Pro Glu Leu Arg Glu Gly Gln Ala Val Val Leu Ser Cys
900 905 910
Gln Val Pro Thr Gly Val Ser Glu Gly Thr Ser Tyr Ser Trp Tyr Gln
915 920 925
Asp Gly Arg Pro Leu Gln Glu Ser Thr Ser Ser Thr Leu Arg Ile Ala
930 935 940
Ala Ile Ser Leu Arg Gln Ala Gly Ala Tyr His Cys Gln Ala Gln Ala
945 950 955 960
Pro Asp Thr Ala Ile Ala Ser Leu Ala Ala Pro Val Ser Leu His Val
965 970 975
Ser Tyr Thr Pro Arg His Val Thr Leu Ser Ala Leu Leu Ser Thr Asp
980 985 990
Pro Glu Arg Leu Gly His Leu Val Cys Ser Val Gln Ser Asp Pro Pro
995 1000 1005
Ala Gln Leu Gln Leu Phe His Arg Asn Arg Leu Val Ala Ser Thr
1010 1015 1020
Leu Gln Gly Ala Asp Glu Leu Ala Gly Ser Asn Pro Arg Leu His
1025 1030 1035
Val Thr Val Leu Pro Asn Glu Leu Arg Leu Gln Ile His Phe Pro
1040 1045 1050
Glu Leu Glu Asp Asp Gly Thr Tyr Thr Cys Glu Ala Ser Asn Thr
1055 1060 1065
Leu Gly Gln Ala Ser Ala Ala Ala Asp Phe Asp Ala Gln Ala Val
1070 1075 1080
Arg Val Thr Val Trp Pro Asn Ala Thr Val Gln Glu Gly Gln Gln
1085 1090 1095
Val Asn Leu Thr Cys Leu Val Trp Ser Thr His Gln Asp Ser Leu
1100 1105 1110
Ser Tyr Thr Trp Tyr Lys Gly Gly Gln Gln Leu Leu Gly Ala Arg
1115 1120 1125
Ser Ile Thr Leu Pro Ser Val Lys Val Leu Asp Ala Thr Ser Tyr
1130 1135 1140
Arg Cys Gly Val Gly Leu Pro Gly His Ala Pro His Leu Ser Arg
1145 1150 1155
Pro Val Thr Leu Asp Val Leu His Ala Pro Arg Asn Leu Arg Leu
1160 1165 1170
Thr Tyr Leu Leu Glu Thr Gln Gly Arg Gln Leu Ala Leu Val Leu
1175 1180 1185
Cys Thr Val Asp Ser Arg Pro Pro Ala Gln Leu Thr Leu Ser His
1190 1195 1200
Gly Asp Gln Leu Val Ala Ser Ser Thr Glu Ala Ser Val Pro Asn
1205 1210 1215
Thr Leu Arg Leu Glu Leu Gln Asp Pro Arg Pro Ser Asn Glu Gly
1220 1225 1230
Leu Tyr Ser Cys Ser Ala His Ser Pro Leu Gly Lys Ala Asn Thr
1235 1240 1245
Ser Leu Glu Leu Leu Leu Glu Gly Val Arg Val Lys Met Asn Pro
1250 1255 1260
Ser Gly Ser Val Pro Glu Gly Glu Pro Val Thr Val Thr Cys Glu
1265 1270 1275
Asp Pro Ala Ala Leu Ser Ser Ala Leu Tyr Ala Trp Phe His Asn
1280 1285 1290
Gly His Trp Leu Gln Glu Gly Pro Ala Ser Ser Leu Gln Phe Leu
1295 1300 1305
Val Thr Thr Arg Ala His Ala Gly Ala Tyr Phe Cys Gln Val His
1310 1315 1320
Asp Thr Gln Gly Thr Arg Ser Ser Arg Pro Ala Ser Leu Gln Ile
1325 1330 1335
Leu Tyr Ala Pro Arg Asp Ala Val Leu Ser Ser Phe Arg Asp Ser
1340 1345 1350
Arg Thr Arg Leu Met Val Val Ile Gln Cys Thr Val Asp Ser Glu
1355 1360 1365
Pro Pro Ala Glu Met Val Leu Ser His Asn Gly Lys Val Leu Ala
1370 1375 1380
Ala Ser His Glu Arg His Ser Ser Ala Ser Gly Ile Gly His Ile
1385 1390 1395
Gln Val Ala Arg Asn Ala Leu Arg Leu Gln Val Gln Asp Val Thr
1400 1405 1410
Leu Gly Asp Gly Asn Thr Tyr Val Cys Thr Ala Gln Asn Thr Leu
1415 1420 1425
Gly Ser Ile Ser Thr Thr Gln Arg Leu Leu Thr Glu Thr Asp Ile
1430 1435 1440
Arg Val Thr Ala Glu Pro Gly Leu Asp Val Pro Glu Gly Thr Ala
1445 1450 1455
Leu Asn Leu Ser Cys Leu Leu Pro Gly Gly Ser Gly Pro Thr Gly
1460 1465 1470
Asn Ser Ser Phe Thr Trp Phe Trp Asn Arg His Arg Leu His Ser
1475 1480 1485
Ala Pro Val Pro Thr Leu Ser Phe Thr Pro Val Val Arg Ala Gln
1490 1495 1500
Ala Gly Leu Tyr His Cys Arg Ala Asp Leu Pro Thr Gly Ala Thr
1505 1510 1515
Thr Ser Ala Pro Val Met Leu Arg Val Leu Tyr Pro Pro Lys Thr
1520 1525 1530
Pro Thr Leu Ile Val Phe Val Glu Pro Gln Gly Gly His Gln Gly
1535 1540 1545
Ile Leu Asp Cys Arg Val Asp Ser Glu Pro Leu Ala Ile Leu Thr
1550 1555 1560
Leu His Arg Gly Ser Gln Leu Val Ala Ser Asn Gln Leu His Asp
1565 1570 1575
Ala Pro Thr Lys Pro His Ile Arg Val Thr Ala Pro Pro Asn Ala
1580 1585 1590
Leu Arg Val Asp Ile Glu Glu Leu Gly Pro Ser Asn Gln Gly Glu
1595 1600 1605
Tyr Val Cys Thr Ala Ser Asn Thr Leu Gly Ser Ala Ser Ala Ser
1610 1615 1620
Ala Tyr Phe Gly Thr Arg Ala Leu His Gln Leu Gln Leu Phe Gln
1625 1630 1635
Arg Leu Leu Trp Val Leu Gly Phe Leu Ala Gly Phe Leu Cys Leu
1640 1645 1650
Leu Leu Gly Leu Val Ala Tyr His Thr Trp Arg Lys Lys Ser Ser
1655 1660 1665
Thr Lys Leu Asn Glu Asp Glu Asn Ser Ala Glu Met Ala Thr Lys
1670 1675 1680
Lys Asn Thr Ile Gln Glu Glu Val Val Ala Ala Leu
1685 1690 1695
<210> SEQ ID NO 13
<211> LENGTH: 6736
<212> TYPE: DNA
<213> ORGANISM: homo sapiens
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/NM_023068
<309> DATABASE ENTRY DATE: 2001-02-13
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(6736)
<400> SEQUENCE: 13
atgggcttct tgcccaagct tctcctcctg gcctcattct tcccagcagg ccaggcctca 60
tggggcgtct ccagtcccca ggacgtgcag ggtgtgaagg ggtcttgcct gcttatcccc 120
tgcatcttca gcttccctgc cgacgtggag gtgcccgacg gcatcacggc catctggtac 180
tacgactact cgggccagcg gcaggtggtg agccactcgg cggaccccaa gctggtggag 240
gcccgcttcc gcggccgcac cgagttcatg gggaaccccg agcacagggt gtgcaacctg 300
ctgctgaagg acctgcagcc cgaggactct ggttcctaca acttccgctt cgagatcagt 360
gaggtcaacc gctggtcaga tgtgaaaggc accttggtca cagtaacaga ggagcccagg 420
gtgcccacca ttgcctcccc ggtggagctt ctcgagggca cagaggtgga cttcaactgc 480
tccactccct acgtatgcct gcaggagcag gtcagactgc agtggcaagg ccaggaccct 540
gctcgctctg tcaccttcaa cagccagaag tttgagccca ccggcgtcgg ccacctggag 600
accctccaca tggccatgtc ctggcaggac cacggccgga tcctgcgctg ccagctctcc 660
gtggccaatc acagggctca gagcgagatt cacctccaag tgaagtatgc ccccaagggt 720
gtgaagatcc tcctcagccc ctcggggagg aacatccttc caggtgagct ggtcacactc 780
acctgccagg tgaacagcag ctaccctgca gtcagttcca ttaagtggct caaggatggg 840
gtacgcctcc aaaccaagac tggtgtgctg cacctgcccc aggcagcctg gagcgatgct 900
ggcgtctaca cctgccaagc tgagaacggc gtgggctctt tggtctcacc ccccatcagc 960
ctccacatct tcatggctga ggtccaggtg agcccagcag gtcccatcct ggagaaccag 1020
acagtgacac tagtctgcaa cacacccaat gaggcaccca gtgatctccg ctacagctgg 1080
tacaagaacc atgtcctgct ggaggatgcc cactcccata ccctccggct gcacttggcc 1140
actagggctg atactggctt ctacttctgt gaggtgcaga acgtccatgg cagcgagcgc 1200
tcgggccctg tcagcgtggt agtcaaccac ccgcctctca ctccagtcct gacagccttc 1260
ctggagaccc aggcgggact tgtgggcatc cttcactgct ctgtggtcag tgagcccctg 1320
gccacactgg tgctgtcaca tgggggtcat atcctggcct ccacctccgg ggacagtgat 1380
cacagcccac gcttcagtgg tacctctggt cccaactccc tgcgcctgga gatccgagac 1440
ctggaggaaa ctgacagtgg ggagtacaag tgctcagcca ccaactccct tggaaatgca 1500
acctccaccc tggacttcca tgccaatgcc gcccgtctcc tcatcagccc ggcagccgag 1560
gtggtggaag gacaggcagt gacactgagc tgcagaagcg gcctaagccc cacacctgat 1620
gcccgcttct cctggtacct gaatggagcc ctgcttcacg agggtcccgg cagcagcctc 1680
ctgctccccg cggcctccag cactgacgcc ggctcatacc actgccgggc ccgggacggc 1740
cacagtgcca gtggcccctc ttcgccagct gttctcactg tgctctaccc ccctcgacaa 1800
ccaacattca ccaccaggct ggaccttgat gccgctgggg ccggggctgg acggcgaggc 1860
ctccttttgt gccgtgtgga cagcgacccc cccgccaggc tgcagctgct ccacaaggac 1920
cgtgttgtgg ccacttccct gccatcaggg ggtggctgca gcacctgtgg gggctgttcc 1980
ccacgcatga aggtcaccaa agcccccaac ttgctgcgtg tggagattca caaccctttg 2040
ctggaagagg agggcttgta cctctgtgag gccagcaatg ccctgggcaa cgcctccacc 2100
tcagccacct tcaatggcca ggccactgtc ctggccattg caccatcaca cacacttcag 2160
gagggcacag aagccaactt gacttgcaac gtgagccggg aagctgctgg cagccctgct 2220
aacttctcct ggttccgaaa tggggtgctg tgggcccagg gtcccctgga gaccgtgaca 2280
ctgctgcccg tggccagaac tgatgctgcc ctttacgcct gccgcatcct gactgaggct 2340
ggtgcccagc tctccactcc cgtgctcctg agtgtactct atcccccgga ccgtccaaag 2400
ctgtcagccc tcctagacat gggccagggc cacatggctc tgttcatctg cactgtggac 2460
agccgccccc tggccttgct ggccttgttc catggggagc acctcctggc caccagcctg 2520
ggtccccagg tcccatccca tggtcggttc caggctaaag ctgaggccaa ctccctgaag 2580
ttagaggtcc gagaactggg ccttggggac tctggcagct accgctgtga ggccacaaat 2640
gttcttggat catccaacac ctcactcttc ttccaggtcc gaggagcctg ggtccaggtg 2700
tcaccatcac ctgagctcca agagggccag gctgtggtcc tgagctgcca ggtacacaca 2760
ggagtcccag aggggacctc atatcgttgg tatcgggatg gccagcccct ccaggagtcg 2820
acctcggcca cgctccgctt tgcagccata actttgacac aagctggggc ctatcattgc 2880
caagcccagg ccccaggctc agccaccacg agcctagctg cacccatcag cctccacgtg 2940
tcctatgccc cacgccacgt cacactcact accctgatgg acacaggccc tggacgactg 3000
ggcctcctcc tgtgccgtgt ggacagtgac cctccggccc agctgcggct gctccacggg 3060
gatcgccttg tggcctccac cctacaaggt gtggggggac ccgaaggcag ctctcccagg 3120
ctgcatgtgg ctgtggcccc caacacactg cgtctggaga tccacggggc tatgctggag 3180
gatgagggtg tctatatctg tgaggcctcc aacaccctgg gccaggcctc ggcctcagct 3240
gacttcgacg ctcaagctgt gaatgtgcag gtgtggcccg gggctaccgt gcgggagggg 3300
cagctggtga acctgacctg ccttgtgtgg accactcacc cggcccagct cacctacaca 3360
tggtaccagg atgggcagca gcgcctggat gcccactcca tccccctgcc caacgtcaca 3420
gtcagggatg ccacctccta ccgctgcggt gtgggccccc ctggtcgggc accccgcctc 3480
tccagaccta tcaccttgga cgtcctctac gcgccccgca acctgcgcct gacctacctc 3540
ctggagagcc atggcgggca gctggccctg gtactgtgca ctgtggacag ccgcccgccc 3600
gcccagctgg ccctcagcca cgccggtcgc ctcttggcct cctcgacagc agcctctgtc 3660
cccaacaccc tgcgcctgga gctgcgaggg ccacagccca gggatgaggg tttctacagc 3720
tgctctgccc gcagccctct gggccaggcc aacacgtccc tggagctgcg gctggagggt 3780
gtgcgggtga tcctggctcc ggaggctgcc gtgcctgaag gtgcccccat cacagtgacc 3840
tgtgcggacc ctgctgccca cgcacccaca ctctatactt ggtaccacaa cggtcgttgg 3900
ctgcaggagg gtccagctgc ctcactctca ttcctggtgg ccacgcgggc tcatgcaggc 3960
gcctactctt gccaggccca ggatgcccag ggcacccgca gctcccgtcc tgctgccctg 4020
caagtcctct atgcccctca ggacgctgtc ctgtcctcct tccgggactc cagggccaga 4080
tccatggctg tgatacagtg cactgtggac agtgagccac ctgctgagct ggccctatct 4140
catgatggca aggtgctggc cacgagcagc ggggtccaca gcttggcatc agggacaggc 4200
catgtccagg tggcccgaaa cgccctacgg ctgcaggtgc aagatgtgcc tgcaggtgat 4260
gacacctatg tttgcacagc ccaaaacttg ctgggctcaa tcagcaccat cgggcggttg 4320
caggtagaag gtgcacgcgt ggtggcagag cctggcctgg acgtgcctga gggcgctgcc 4380
ctgaacctca gctgccgcct cctgggtggc cctgggcctg tgggcaactc cacctttgca 4440
tggttctgga atgaccggcg gctgcacgcg gagcctgtgc ccactctcgc cttcacccac 4500
gtggctcgtg ctcaagctgg gatgtaccac tgcctggctg agctccccac tggggctgct 4560
gcctctgctc cagtcatgct ccgtgtgctc taccctccca agacgcccac catgatggtc 4620
ttcgtggagc ctgagggtgg cctccggggc atcctggatt gccgagtgga cagcgagccg 4680
ctcgccagcc tgactctcca ccttggcagt cgactggtgg cctccagtca gccccagggt 4740
gctcctgcag agccacacat ccatgtcctg gcttccccca atgccctgag ggtggacatc 4800
gaggcgctga ggcccagcga ccaaggggaa tacatctgtt ctgcctcaaa tgtcctgggc 4860
tctgcctcta cctccaccta ctttggggtc agagccctgc accgcctgca tcagttccag 4920
cagctgctct gggtcctggg actgctggtg ggcctcctgc tcctgctgtt gggcctgggg 4980
gcctgctaca cctggagaag gaggcgtgtt tgtaagcaga gcatgggcga gaattcggtg 5040
gagatggctt ttcagaaaga gaccacgcag ctcattgatc ctgatgcagc cacatgtgag 5100
acctcaacct gtgccccacc cctgggctga ccagtggtgt tgcctgccct ccggaggaga 5160
aagtggccag aatctgtgat gactccagcc tatgaatgtg aatgaggcag tgttgagtcc 5220
tgcccgcctc tacgaaaaca gctctgtgac atctgacttt ttatgacctg gccccaagcc 5280
tcttgccccc ccaaaaatgg gtggtgagag gtctgcccag gagggtgttg accctggagg 5340
acactgaaga gcactgagct gatctcgctc tctcttctct ggatctcctc ccttctctcc 5400
atttctccct caaaggaagc cctgcccttt cacatccttc tcctcgaaag tcaccctgga 5460
ctttggttgg attgcagcat cctgcatcct cagaggctca ccaaggcatt ctgtattcaa 5520
cagagtatca gtcagcctgc tctaacaaga gaccaaatac agtgacttca acatgataga 5580
attttatttt tctctcccac gctagtctgg ctgttacgat ggtttatgat gttggggctc 5640
aggatccttc tatcttcctt ttctctatcc ctaaaatgat gcctttgatt gtgaggctca 5700
ccatggcccc gctttgtcca catgccctcc agccagaaga aggaagagtg gaggtagaag 5760
cacacccatg cccatggtgg acgcaactca gaagctgcac aggacttttc cactcacttc 5820
ccattggctg gagtattgtc acatggctac tgcaagctac aagggagact gggaaatgta 5880
gtttttattt tgagtccaga ggacatttgg aattggactt ccaaaggact cccaactgtg 5940
agctcatccc tgagactttt gacattgttg ggaatgccac cagcaggcca tgttttgtct 6000
cagtgcccat ctactgaggg ccagggtgtg cccctggcca ttctggttgt gggcttcctg 6060
gaagaggtga tcactctcac actaagactg aggaaataaa aaaggtttgg tgttttccta 6120
gggagagagc atgccaggca gtggagttgc ctaagcagac atccttgtgc cagatttggc 6180
ccctgaaaga agagatgccc tcattcccac caccaccccc cctaccccca gggactgggt 6240
actaccttac tggcccttac aagagtggag ggcagacaca gatgttgtca gcatccttat 6300
tcctgctcca gatgcatctc tgttcatgac tgtgtgagct cctgtccttt tcctggagac 6360
cctgtgtcgg gctgttaaag agaatgagtt accaagaagg aatgacgtgc ccctgcgaat 6420
cagggaccaa caggagagag ctcttgagtg ggctagtgac tccccctgca gcctggtgga 6480
gatggtgtga ggagcgaaga gccctctgct ctaggatttg ggttgaaaaa cagagagaga 6540
agtggggagt tgccacagga gctaacacgc tgggaggcag ttgggggcgg gtgaactttg 6600
tgtagccgag gccgcaccct ccctcattcc aggctcattc attttcatgc tccattgcca 6660
gactcttgct gggagcccgt ccagaatgtc ctcccaataa aactccatcc tatgacgcaa 6720
aaaaaaaaaa aaaaaa 6736
<210> SEQ ID NO 14
<211> LENGTH: 1709
<212> TYPE: PRT
<213> ORGANISM: homo sapiens
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/NP_075556
<309> DATABASE ENTRY DATE: 2001-02-13
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1709)
<400> SEQUENCE: 14
Met Gly Phe Leu Pro Lys Leu Leu Leu Leu Ala Ser Phe Phe Pro Ala
1 5 10 15
Gly Gln Ala Ser Trp Gly Val Ser Ser Pro Gln Asp Val Gln Gly Val
20 25 30
Lys Gly Ser Cys Leu Leu Ile Pro Cys Ile Phe Ser Phe Pro Ala Asp
35 40 45
Val Glu Val Pro Asp Gly Ile Thr Ala Ile Trp Tyr Tyr Asp Tyr Ser
50 55 60
Gly Gln Arg Gln Val Val Ser His Ser Ala Asp Pro Lys Leu Val Glu
65 70 75 80
Ala Arg Phe Arg Gly Arg Thr Glu Phe Met Gly Asn Pro Glu His Arg
85 90 95
Val Cys Asn Leu Leu Leu Lys Asp Leu Gln Pro Glu Asp Ser Gly Ser
100 105 110
Tyr Asn Phe Arg Phe Glu Ile Ser Glu Val Asn Arg Trp Ser Asp Val
115 120 125
Lys Gly Thr Leu Val Thr Val Thr Glu Glu Pro Arg Val Pro Thr Ile
130 135 140
Ala Ser Pro Val Glu Leu Leu Glu Gly Thr Glu Val Asp Phe Asn Cys
145 150 155 160
Ser Thr Pro Tyr Val Cys Leu Gln Glu Gln Val Arg Leu Gln Trp Gln
165 170 175
Gly Gln Asp Pro Ala Arg Ser Val Thr Phe Asn Ser Gln Lys Phe Glu
180 185 190
Pro Thr Gly Val Gly His Leu Glu Thr Leu His Met Ala Met Ser Trp
195 200 205
Gln Asp His Gly Arg Ile Leu Arg Cys Gln Leu Ser Val Ala Asn His
210 215 220
Arg Ala Gln Ser Glu Ile His Leu Gln Val Lys Tyr Ala Pro Lys Gly
225 230 235 240
Val Lys Ile Leu Leu Ser Pro Ser Gly Arg Asn Ile Leu Pro Gly Glu
245 250 255
Leu Val Thr Leu Thr Cys Gln Val Asn Ser Ser Tyr Pro Ala Val Ser
260 265 270
Ser Ile Lys Trp Leu Lys Asp Gly Val Arg Leu Gln Thr Lys Thr Gly
275 280 285
Val Leu His Leu Pro Gln Ala Ala Trp Ser Asp Ala Gly Val Tyr Thr
290 295 300
Cys Gln Ala Glu Asn Gly Val Gly Ser Leu Val Ser Pro Pro Ile Ser
305 310 315 320
Leu His Ile Phe Met Ala Glu Val Gln Val Ser Pro Ala Gly Pro Ile
325 330 335
Leu Glu Asn Gln Thr Val Thr Leu Val Cys Asn Thr Pro Asn Glu Ala
340 345 350
Pro Ser Asp Leu Arg Tyr Ser Trp Tyr Lys Asn His Val Leu Leu Glu
355 360 365
Asp Ala His Ser His Thr Leu Arg Leu His Leu Ala Thr Arg Ala Asp
370 375 380
Thr Gly Phe Tyr Phe Cys Glu Val Gln Asn Val His Gly Ser Glu Arg
385 390 395 400
Ser Gly Pro Val Ser Val Val Val Asn His Pro Pro Leu Thr Pro Val
405 410 415
Leu Thr Ala Phe Leu Glu Thr Gln Ala Gly Leu Val Gly Ile Leu His
420 425 430
Cys Ser Val Val Ser Glu Pro Leu Ala Thr Leu Val Leu Ser His Gly
435 440 445
Gly His Ile Leu Ala Ser Thr Ser Gly Asp Ser Asp His Ser Pro Arg
450 455 460
Phe Ser Gly Thr Ser Gly Pro Asn Ser Leu Arg Leu Glu Ile Arg Asp
465 470 475 480
Leu Glu Glu Thr Asp Ser Gly Glu Tyr Lys Cys Ser Ala Thr Asn Ser
485 490 495
Leu Gly Asn Ala Thr Ser Thr Leu Asp Phe His Ala Asn Ala Ala Arg
500 505 510
Leu Leu Ile Ser Pro Ala Ala Glu Val Val Glu Gly Gln Ala Val Thr
515 520 525
Leu Ser Cys Arg Ser Gly Leu Ser Pro Thr Pro Asp Ala Arg Phe Ser
530 535 540
Trp Tyr Leu Asn Gly Ala Leu Leu His Glu Gly Pro Gly Ser Ser Leu
545 550 555 560
Leu Leu Pro Ala Ala Ser Ser Thr Asp Ala Gly Ser Tyr His Cys Arg
565 570 575
Ala Arg Asp Gly His Ser Ala Ser Gly Pro Ser Ser Pro Ala Val Leu
580 585 590
Thr Val Leu Tyr Pro Pro Arg Gln Pro Thr Phe Thr Thr Arg Leu Asp
595 600 605
Leu Asp Ala Ala Gly Ala Gly Ala Gly Arg Arg Gly Leu Leu Leu Cys
610 615 620
Arg Val Asp Ser Asp Pro Pro Ala Arg Leu Gln Leu Leu His Lys Asp
625 630 635 640
Arg Val Val Ala Thr Ser Leu Pro Ser Gly Gly Gly Cys Ser Thr Cys
645 650 655
Gly Gly Cys Ser Pro Arg Met Lys Val Thr Lys Ala Pro Asn Leu Leu
660 665 670
Arg Val Glu Ile His Asn Pro Leu Leu Glu Glu Glu Gly Leu Tyr Leu
675 680 685
Cys Glu Ala Ser Asn Ala Leu Gly Asn Ala Ser Thr Ser Ala Thr Phe
690 695 700
Asn Gly Gln Ala Thr Val Leu Ala Ile Ala Pro Ser His Thr Leu Gln
705 710 715 720
Glu Gly Thr Glu Ala Asn Leu Thr Cys Asn Val Ser Arg Glu Ala Ala
725 730 735
Gly Ser Pro Ala Asn Phe Ser Trp Phe Arg Asn Gly Val Leu Trp Ala
740 745 750
Gln Gly Pro Leu Glu Thr Val Thr Leu Leu Pro Val Ala Arg Thr Asp
755 760 765
Ala Ala Leu Tyr Ala Cys Arg Ile Leu Thr Glu Ala Gly Ala Gln Leu
770 775 780
Ser Thr Pro Val Leu Leu Ser Val Leu Tyr Pro Pro Asp Arg Pro Lys
785 790 795 800
Leu Ser Ala Leu Leu Asp Met Gly Gln Gly His Met Ala Leu Phe Ile
805 810 815
Cys Thr Val Asp Ser Arg Pro Leu Ala Leu Leu Ala Leu Phe His Gly
820 825 830
Glu His Leu Leu Ala Thr Ser Leu Gly Pro Gln Val Pro Ser His Gly
835 840 845
Arg Phe Gln Ala Lys Ala Glu Ala Asn Ser Leu Lys Leu Glu Val Arg
850 855 860
Glu Leu Gly Leu Gly Asp Ser Gly Ser Tyr Arg Cys Glu Ala Thr Asn
865 870 875 880
Val Leu Gly Ser Ser Asn Thr Ser Leu Phe Phe Gln Val Arg Gly Ala
885 890 895
Trp Val Gln Val Ser Pro Ser Pro Glu Leu Gln Glu Gly Gln Ala Val
900 905 910
Val Leu Ser Cys Gln Val His Thr Gly Val Pro Glu Gly Thr Ser Tyr
915 920 925
Arg Trp Tyr Arg Asp Gly Gln Pro Leu Gln Glu Ser Thr Ser Ala Thr
930 935 940
Leu Arg Phe Ala Ala Ile Thr Leu Thr Gln Ala Gly Ala Tyr His Cys
945 950 955 960
Gln Ala Gln Ala Pro Gly Ser Ala Thr Thr Ser Leu Ala Ala Pro Ile
965 970 975
Ser Leu His Val Ser Tyr Ala Pro Arg His Val Thr Leu Thr Thr Leu
980 985 990
Met Asp Thr Gly Pro Gly Arg Leu Gly Leu Leu Leu Cys Arg Val Asp
995 1000 1005
Ser Asp Pro Pro Ala Gln Leu Arg Leu Leu His Gly Asp Arg Leu
1010 1015 1020
Val Ala Ser Thr Leu Gln Gly Val Gly Gly Pro Glu Gly Ser Ser
1025 1030 1035
Pro Arg Leu His Val Ala Val Ala Pro Asn Thr Leu Arg Leu Glu
1040 1045 1050
Ile His Gly Ala Met Leu Glu Asp Glu Gly Val Tyr Ile Cys Glu
1055 1060 1065
Ala Ser Asn Thr Leu Gly Gln Ala Ser Ala Ser Ala Asp Phe Asp
1070 1075 1080
Ala Gln Ala Val Asn Val Gln Val Trp Pro Gly Ala Thr Val Arg
1085 1090 1095
Glu Gly Gln Leu Val Asn Leu Thr Cys Leu Val Trp Thr Thr His
1100 1105 1110
Pro Ala Gln Leu Thr Tyr Thr Trp Tyr Gln Asp Gly Gln Gln Arg
1115 1120 1125
Leu Asp Ala His Ser Ile Pro Leu Pro Asn Val Thr Val Arg Asp
1130 1135 1140
Ala Thr Ser Tyr Arg Cys Gly Val Gly Pro Pro Gly Arg Ala Pro
1145 1150 1155
Arg Leu Ser Arg Pro Ile Thr Leu Asp Val Leu Tyr Ala Pro Arg
1160 1165 1170
Asn Leu Arg Leu Thr Tyr Leu Leu Glu Ser His Gly Gly Gln Leu
1175 1180 1185
Ala Leu Val Leu Cys Thr Val Asp Ser Arg Pro Pro Ala Gln Leu
1190 1195 1200
Ala Leu Ser His Ala Gly Arg Leu Leu Ala Ser Ser Thr Ala Ala
1205 1210 1215
Ser Val Pro Asn Thr Leu Arg Leu Glu Leu Arg Gly Pro Gln Pro
1220 1225 1230
Arg Asp Glu Gly Phe Tyr Ser Cys Ser Ala Arg Ser Pro Leu Gly
1235 1240 1245
Gln Ala Asn Thr Ser Leu Glu Leu Arg Leu Glu Gly Val Arg Val
1250 1255 1260
Ile Leu Ala Pro Glu Ala Ala Val Pro Glu Gly Ala Pro Ile Thr
1265 1270 1275
Val Thr Cys Ala Asp Pro Ala Ala His Ala Pro Thr Leu Tyr Thr
1280 1285 1290
Trp Tyr His Asn Gly Arg Trp Leu Gln Glu Gly Pro Ala Ala Ser
1295 1300 1305
Leu Ser Phe Leu Val Ala Thr Arg Ala His Ala Gly Ala Tyr Ser
1310 1315 1320
Cys Gln Ala Gln Asp Ala Gln Gly Thr Arg Ser Ser Arg Pro Ala
1325 1330 1335
Ala Leu Gln Val Leu Tyr Ala Pro Gln Asp Ala Val Leu Ser Ser
1340 1345 1350
Phe Arg Asp Ser Arg Ala Arg Ser Met Ala Val Ile Gln Cys Thr
1355 1360 1365
Val Asp Ser Glu Pro Pro Ala Glu Leu Ala Leu Ser His Asp Gly
1370 1375 1380
Lys Val Leu Ala Thr Ser Ser Gly Val His Ser Leu Ala Ser Gly
1385 1390 1395
Thr Gly His Val Gln Val Ala Arg Asn Ala Leu Arg Leu Gln Val
1400 1405 1410
Gln Asp Val Pro Ala Gly Asp Asp Thr Tyr Val Cys Thr Ala Gln
1415 1420 1425
Asn Leu Leu Gly Ser Ile Ser Thr Ile Gly Arg Leu Gln Val Glu
1430 1435 1440
Gly Ala Arg Val Val Ala Glu Pro Gly Leu Asp Val Pro Glu Gly
1445 1450 1455
Ala Ala Leu Asn Leu Ser Cys Arg Leu Leu Gly Gly Pro Gly Pro
1460 1465 1470
Val Gly Asn Ser Thr Phe Ala Trp Phe Trp Asn Asp Arg Arg Leu
1475 1480 1485
His Ala Glu Pro Val Pro Thr Leu Ala Phe Thr His Val Ala Arg
1490 1495 1500
Ala Gln Ala Gly Met Tyr His Cys Leu Ala Glu Leu Pro Thr Gly
1505 1510 1515
Ala Ala Ala Ser Ala Pro Val Met Leu Arg Val Leu Tyr Pro Pro
1520 1525 1530
Lys Thr Pro Thr Met Met Val Phe Val Glu Pro Glu Gly Gly Leu
1535 1540 1545
Arg Gly Ile Leu Asp Cys Arg Val Asp Ser Glu Pro Leu Ala Ser
1550 1555 1560
Leu Thr Leu His Leu Gly Ser Arg Leu Val Ala Ser Ser Gln Pro
1565 1570 1575
Gln Gly Ala Pro Ala Glu Pro His Ile His Val Leu Ala Ser Pro
1580 1585 1590
Asn Ala Leu Arg Val Asp Ile Glu Ala Leu Arg Pro Ser Asp Gln
1595 1600 1605
Gly Glu Tyr Ile Cys Ser Ala Ser Asn Val Leu Gly Ser Ala Ser
1610 1615 1620
Thr Ser Thr Tyr Phe Gly Val Arg Ala Leu His Arg Leu His Gln
1625 1630 1635
Phe Gln Gln Leu Leu Trp Val Leu Gly Leu Leu Val Gly Leu Leu
1640 1645 1650
Leu Leu Leu Leu Gly Leu Gly Ala Cys Tyr Thr Trp Arg Arg Arg
1655 1660 1665
Arg Val Cys Lys Gln Ser Met Gly Glu Asn Ser Val Glu Met Ala
1670 1675 1680
Phe Gln Lys Glu Thr Thr Gln Leu Ile Asp Pro Asp Ala Ala Thr
1685 1690 1695
Cys Glu Thr Ser Thr Cys Ala Pro Pro Leu Gly
1700 1705
<210> SEQ ID NO 15
<211> LENGTH: 40
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: PCR forward primer CD163
<400> SEQUENCE: 15
cac cat gga caa act cag aat ggt gct aca tga aaa ctc t 40
<210> SEQ ID NO 16
<211> LENGTH: 33
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: PCR reverse primer CD163
<400> SEQUENCE: 16
tca ttg tac ttc aga gtg gtc tcc tga ggg att 33
1
SEQUENCE LISTING
<160> NUMBER OF SEQ ID NOS: 16
<210> SEQ ID NO 1
<211> LENGTH: 3400
<212> TYPE: DNA
<213> ORGANISM: Sus scrofa
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/AJ311716
<309> DATABASE ENTRY DATE: 2005-04-15
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(3400)
<400> SEQUENCE: 1
atggtgctac ttgaagactc tggatctgca gactttagaa gatgttctgc ccatttaagt 60
tccttcactt ttgctgtagt cgctgttctc agtgcctgct tggtcactag ttctcttgga 120
ggaaaagaca aggagctgag gctaacgggt ggtgaaaaca agtgctctgg aagagtggag 180
gtgaaagtgc aggaggagtg gggaactgtg tgtaataatg gctgggacat ggatgtggtc 240
tctgttgttt gtaggcagct gggatgtcca actgctatca aagccactgg atgggctaat 300
tttagtgcag gttctggacg catttggatg gatcatgttt cttgtcgagg gaatgagtca 360
gctctctggg actgcaaaca tgatggatgg ggaaagcata actgtactca ccaacaggat 420
gctggagtaa cctgctcaga tggatctgat ttagagatga ggctggtgaa tggaggaaac 480
cggtgcttag gaagaataga agtcaaattt caagagcggt ggggaacagt gtgtgatgat 540
aacttcaaca taaatcatgc ttctgtggtt tgtaaacaac ttgaatgtgg aagtgctgtc 600
agtttctctg gttcagctaa ttttggagaa ggttctggac caatctggtt tgatgatctt 660
gtatgcaatg gaaatgagtc agctctctgg aactgcaaac atgaaggatg gggaaagcac 720
aattgcgatc atgctgagga tgctggagtg atttgcttaa atggagcaga cctgaaactg 780
agagtggtag atggactcac tgaatgttca ggaagattgg aagtgaaatt ccaaggagaa 840
tggggaacaa tctgtgatga tggctgggat agtgatgatg ccgctgtggc atgtaagcaa 900
ctgggatgtc caactgctgt cactgccatt ggtcgagtta acgccagtga gggaactgga 960
cacatttggc ttgacagtgt ttcttgccat ggacacgagt ctgctctctg gcagtgtaga 1020
caccatgaat ggggaaagca ttattgcaat cataatgaag atgctggtgt gacatgttct 1080
gatggatcag atctggaact gagacttaaa ggtggaggca gccactgtgc tgggacagtg 1140
gaggtggaaa ttcagaaact ggtaggaaaa gtgtgtgata gaagctgggg actgaaagaa 1200
gctgatgtgg tttgcaggca gctgggatgt ggatctgcac tcaaaacatc atatcaagtt 1260
tattccaaaa ccaaggcaac aaacacatgg ctgtttgtaa gcagctgtaa tggaaatgaa 1320
acttctcttt gggactgcaa gaattggcag tggggtggac ttagttgtga tcactatgac 1380
gaagccaaaa ttacctgctc agcccacagg aaacccaggc tggttggagg ggacattccc 1440
tgctctggtc gtgttgaagt acaacatgga gacacgtggg gcaccgtctg tgattctgac 1500
ttctctctgg aggcggccag cgtgctgtgc agggaactac agtgcggcac tgtggtttcc 1560
ctcctggggg gagctcactt tggagaagga agtggacaga tctgggctga agaattccag 1620
tgtgaggggc acgagtccca cctttcactc tgcccagtag caccccgccc tgacgggaca 1680
tgtagccaca gcagggacgt cggcgtagtc tgctcaagat acacacaaat ccgcttggtg 1740
aatggcaaga ccccatgtga aggaagagtg gagctcaaca ttcttgggtc ctgggggtcc 1800
ctctgcaact ctcactggga catggaagat gcccatgttt tatgccagca gcttaaatgt 1860
ggagttgccc tttctatccc gggaggagca ccttttggga aaggaagtga gcaggtctgg 1920
aggcacatgt ttcactgcac tgggactgag aagcacatgg gagattgttc cgtcactgct 1980
ctgggcgcat cactctgttc ttcagggcaa gtggcctctg taatctgctc agggaaccag 2040
agtcagacac tatccccgtg caattcatca tcctcggacc catcaagctc tattatttca 2100
gaagaaagtg gtgttgcctg catagggagt ggtcaacttc gcctggtcga tggaggtggt 2160
cgttgtgctg ggagagtaga ggtctatcct ggggcatcct ggggcaccat ctgtgatgac 2220
agctgggacc tgaatgatgc ccatgtggtg tgcaaacagc tgagctgtgg atgggccatt 2280
aatgccactg gttctgctca ttttggggaa ggaacagggc ccatttggct ggatgagata 2340
aactgtaatg gaaaagaatc tcatatttgg caatgccact cacatggttg ggggcggcac 2400
aattgcaggc ataaggagga tgcaggagtc atctgctcag agttcatgtc tctgagactg 2460
atcagtgaaa acagcagaga gacctgtgca gggcgcctgg aagtttttta caacggagct 2520
tggggcagcg ttggcaggaa tagcatgtct ccagccacag tgggggtggt atgcaggcag 2580
ctgggctgtg cagacagagg ggacatcagc cctgcatctt cagacaagac agtgtccagg 2640
cacatgtggg tggacaatgt tcagtgtcct aaaggacctg acacactatg gcagtgcccc 2700
tcatctccat ggaagaagag actggccagc ccctcagagg agacatggat cacatgtgcc 2760
aacaaaataa gacttcaaga aggaaacact aattgttctg gacgtgtgga gatctggtac 2820
ggaggttcct ggggcactgt gtgtgacgac tcctgggacc ttgaagatgc tcaggtggtg 2880
tgccgacagc tgggctgtgg ctcagctttg gaggcaggaa aagagcccgc atttggccag 2940
gggactgggc ccatatggct caatgaagtg aagtgcaagg ggaatgaacc ctccttgtgg 3000
gattgtcctg ccagatcctg gggccacagt gactgtggac acaaggagga tgctgctgtg 3060
acgtgctcag aaattgcaaa gagccgagaa tccctacatg ccacaggtcg ctcatctttt 3120
gttgcacttg caatctttgg ggtcattctg ttggcctgtc tcatcgcatt cctcatttgg 3180
actcagaagc gaagacagag gcagcggctc tcagttttct caggaggaga gaattctgtc 3240
catcaaattc aataccggga gatgaattct tgcctgaaag cagatgaaac ggatatgcta 3300
aatccctcag gagaccactc tgaagtacaa tgaaaaggaa aatgggaatt ataacctggt 3360
gagttcagcc tttaagatac cttgatgaag acctggacta 3400
<210> SEQ ID NO 2
<211> LENGTH: 1115
<212> TYPE: PRT
<213> ORGANISM: Sus scrofa
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/CAC84397
<309> DATABASE ENTRY DATE: 2005-04-15
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1115)
<400> SEQUENCE: 2
Met Asp Lys Leu Arg Met Val Leu His Glu Asn Ser Gly Ser Ala Asp
1 5 10 15
Phe Arg Arg Cys Ser Ala His Leu Ser Ser Phe Thr Phe Ala Val Val
20 25 30
Ala Val Leu Ser Ala Cys Leu Val Thr Ser Ser Leu Gly Gly Lys Asp
35 40 45
Lys Glu Leu Arg Leu Thr Gly Gly Glu Asn Lys Cys Ser Gly Arg Val
50 55 60
Glu Val Lys Val Gln Glu Glu Trp Gly Thr Val Cys Asn Asn Gly Trp
65 70 75 80
Asp Met Asp Val Val Ser Val Val Cys Arg Gln Leu Gly Cys Pro Thr
85 90 95
Ala Ile Lys Ala Thr Gly Trp Ala Asn Phe Ser Ala Gly Ser Gly Arg
100 105 110
Ile Trp Met Asp His Val Ser Cys Arg Gly Asn Glu Ser Ala Leu Trp
115 120 125
Asp Cys Lys His Asp Gly Trp Gly Lys His Asn Cys Thr His Gln Gln
130 135 140
Asp Ala Gly Val Thr Cys Ser Asp Gly Ser Asp Leu Glu Met Gly Leu
145 150 155 160
Val Asn Gly Gly Asn Arg Cys Leu Gly Arg Ile Glu Val Lys Phe Gln
165 170 175
Gly Arg Trp Gly Thr Val Cys Asp Asp Asn Phe Asn Ile Asn His Ala
180 185 190
Ser Val Val Cys Lys Gln Leu Glu Cys Gly Ser Ala Val Ser Phe Ser
195 200 205
Gly Ser Ala Asn Phe Gly Glu Gly Ser Gly Pro Ile Trp Phe Asp Asp
210 215 220
Leu Val Cys Asn Gly Asn Glu Ser Ala Leu Trp Asn Cys Lys His Glu
225 230 235 240
Gly Trp Gly Lys His Asn Cys Asp His Ala Glu Asp Ala Gly Val Ile
245 250 255
Cys Leu Asn Gly Ala Asp Leu Lys Leu Arg Val Val Asp Gly Val Thr
260 265 270
Glu Cys Ser Gly Arg Leu Glu Val Lys Phe Gln Gly Glu Trp Gly Thr
275 280 285
Ile Cys Asp Asp Gly Trp Asp Ser Asp Asp Ala Ala Val Ala Cys Lys
290 295 300
Gln Leu Gly Cys Pro Thr Ala Val Thr Ala Ile Gly Arg Val Asn Ala
305 310 315 320
Ser Glu Gly Thr Gly His Ile Trp Leu Asp Ser Val Ser Cys His Gly
325 330 335
His Glu Ser Ala Leu Trp Gln Cys Arg His His Glu Trp Gly Lys His
340 345 350
Tyr Cys Asn His Asp Glu Asp Ala Gly Val Thr Cys Ser Asp Gly Ser
355 360 365
Asp Leu Glu Leu Arg Leu Lys Gly Gly Gly Ser His Cys Ala Gly Thr
370 375 380
Val Glu Val Glu Ile Gln Lys Leu Val Gly Lys Val Cys Asp Arg Ser
385 390 395 400
Trp Gly Leu Lys Glu Ala Asp Val Val Cys Arg Gln Leu Gly Cys Gly
405 410 415
Ser Ala Leu Lys Thr Ser Tyr Gln Val Tyr Ser Lys Thr Lys Ala Thr
420 425 430
Asn Thr Trp Leu Phe Val Ser Ser Cys Asn Gly Asn Glu Thr Ser Leu
435 440 445
Trp Asp Cys Lys Asn Trp Gln Trp Gly Gly Leu Ser Cys Asp His Tyr
450 455 460
Asp Glu Ala Lys Ile Thr Cys Ser Ala His Arg Lys Pro Arg Leu Val
465 470 475 480
Gly Gly Asp Ile Pro Cys Ser Gly Arg Val Glu Val Gln His Gly Asp
485 490 495
Thr Trp Gly Thr Val Cys Asp Ser Asp Phe Ser Leu Glu Ala Ala Ser
500 505 510
Val Leu Cys Arg Glu Leu Gln Cys Gly Thr Val Val Ser Leu Leu Gly
515 520 525
Gly Ala His Phe Gly Glu Gly Ser Gly Gln Ile Trp Ala Glu Glu Phe
530 535 540
Gln Cys Glu Gly His Glu Ser His Leu Ser Leu Cys Pro Val Ala Pro
545 550 555 560
Arg Pro Asp Gly Thr Cys Ser His Ser Arg Asp Val Gly Val Val Cys
565 570 575
Ser Arg Tyr Thr Gln Ile Arg Leu Val Asn Gly Lys Thr Pro Cys Glu
580 585 590
Gly Arg Val Glu Leu Asn Ile Leu Gly Ser Trp Gly Ser Leu Cys Asn
595 600 605
Ser His Trp Asp Met Glu Asp Ala His Val Leu Cys Gln Gln Leu Lys
610 615 620
Cys Gly Val Ala Leu Ser Ile Pro Gly Gly Ala Pro Phe Gly Lys Gly
625 630 635 640
Ser Glu Gln Val Trp Arg His Met Phe His Cys Thr Gly Thr Glu Lys
645 650 655
His Met Gly Asp Cys Ser Val Thr Ala Leu Gly Ala Ser Leu Cys Ser
660 665 670
Ser Gly Gln Val Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln Thr
675 680 685
Leu Ser Pro Cys Asn Ser Ser Ser Ser Asp Pro Ser Ser Ser Ile Ile
690 695 700
Ser Glu Glu Asn Gly Val Ala Cys Ile Gly Ser Gly Gln Leu Arg Leu
705 710 715 720
Val Asp Gly Gly Gly Arg Cys Ala Gly Arg Val Glu Val Tyr His Glu
725 730 735
Gly Ser Trp Gly Thr Ile Cys Asp Asp Ser Trp Asp Leu Asn Asp Ala
740 745 750
His Val Val Cys Lys Gln Leu Ser Cys Gly Trp Ala Ile Asn Ala Thr
755 760 765
Gly Ser Ala His Phe Gly Glu Gly Thr Gly Pro Ile Trp Leu Asp Glu
770 775 780
Ile Asn Cys Asn Gly Lys Glu Ser His Ile Trp Gln Cys His Ser His
785 790 795 800
Gly Trp Gly Arg His Asn Cys Arg His Lys Glu Asp Ala Gly Val Ile
805 810 815
Cys Ser Glu Phe Met Ser Leu Arg Leu Ile Ser Glu Asn Ser Arg Glu
820 825 830
Thr Cys Ala Gly Arg Leu Glu Val Phe Tyr Asn Gly Ala Trp Gly Ser
835 840 845
Val Gly Arg Asn Ser Met Ser Pro Ala Thr Val Gly Val Val Cys Arg
850 855 860
Gln Leu Gly Cys Ala Asp Arg Gly Asp Ile Ser Pro Ala Ser Ser Asp
865 870 875 880
Lys Thr Val Ser Arg His Met Trp Val Asp Asn Val Gln Cys Pro Lys
885 890 895
Gly Pro Asp Thr Leu Trp Gln Cys Pro Ser Ser Pro Trp Lys Lys Arg
900 905 910
Leu Ala Ser Pro Ser Glu Glu Thr Trp Ile Thr Cys Ala Asn Lys Ile
915 920 925
Arg Leu Gln Glu Gly Asn Thr Asn Cys Ser Gly Arg Val Glu Ile Trp
930 935 940
Tyr Gly Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu Glu
945 950 955 960
Asp Ala Gln Val Val Cys Arg Gln Leu Gly Cys Gly Ser Ala Leu Glu
965 970 975
Ala Gly Lys Glu Ala Ala Phe Gly Gln Gly Thr Gly Pro Ile Trp Leu
980 985 990
Asn Glu Val Lys Cys Lys Gly Asn Glu Thr Ser Leu Trp Asp Cys Pro
995 1000 1005
Ala Arg Ser Trp Gly His Ser Asp Cys Gly His Lys Glu Asp Ala
1010 1015 1020
Ala Val Thr Cys Ser Glu Ile Ala Lys Ser Arg Glu Ser Leu His
1025 1030 1035
Ala Thr Gly Arg Ser Ser Phe Val Ala Leu Ala Ile Phe Gly Val
1040 1045 1050
Ile Leu Leu Ala Cys Leu Ile Ala Phe Leu Ile Trp Thr Gln Lys
1055 1060 1065
Arg Arg Gln Arg Gln Arg Leu Ser Val Phe Ser Gly Gly Glu Asn
1070 1075 1080
Ser Val His Gln Ile Gln Tyr Arg Glu Met Asn Ser Cys Leu Lys
1085 1090 1095
Ala Asp Glu Thr Asp Met Leu Asn Pro Ser Gly Asp His Ser Glu
1100 1105 1110
Val Gln
1115
<210> SEQ ID NO 3
<211> LENGTH: 4405
<212> TYPE: DNA
<213> ORGANISM: mus musculus
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/AF274883
<309> DATABASE ENTRY DATE: 2001-05-10
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(4405)
<400> SEQUENCE: 3
gtggtcatcc actttctaca gagaacacgt ctatgaaata gtatcaggag acacacggag 60
ccatcaaaat catcaagctt tggaatgggt ggacacagaa tggttcttct tggaggtgct 120
ggatctcctg gttgtaaaag gtttgtccat ctaggtttct ttgttgtggc tgtgagctca 180
cttctcagtg cctctgctgt cactaacgct cctggagaaa tgaagaagga actgagactg 240
gcgggtggtg aaaacaactg tagtgggaga gtggaactta agatccatga caagtggggc 300
acagtgtgca gtaacggctg gagcatgaat gaagtgtccg tggtttgcca gcagctggga 360
tgcccaactt ctattaaagc ccttggatgg gctaactcca gcgccggctc tggatatatc 420
tggatggaca aagtttcttg tacagggaat gagtcagctc tttgggactg caaacatgat 480
gggtggggaa agcataactg tacccatgaa aaagatgctg gagtgacctg ctcagatgga 540
tctaatttgg agatgagact ggtgaacagt gcgggccacc gatgcttagg aagagtagaa 600
ataaagttcc agggaaagtg ggggacggtg tgtgacgaca acttcagcaa agatcacgct 660
tctgtgattt gtaaacagct tggatgtgga agtgccatta gtttctctgg ctcagctaaa 720
ttgggagctg gttctggacc aatctggctc gatgacctgg catgcaatgg aaatgagtca 780
gctctctggg actgcaaaca ccggggatgg ggcaagcata actgtgacca tgctgaggat 840
gtcggtgtga tttgcttaga gggagcagat ctgagcctga gactagtgga tggagtgtcc 900
agatgttcag gaagattgga agtgagattc caaggagaat gggggaccgt gtgtgatgat 960
aactgggatc tccgggatgc ttctgtggtg tgcaagcaac tgggatgtcc aactgccatc 1020
agtgccattg gtcgagttaa tgccagtgag ggatctggac agatttggct tgacaacatt 1080
tcatgcgaag gacatgaggc aactctttgg gagtgtaaac accaagagtg gggaaagcat 1140
tactgtcatc atagagaaga cgctggtgtg acatgttctg atggagcaga tctggaactt 1200
agacttgtag gtggaggcag tcgctgtgct ggcattgtgg aggtggagat tcagaagctg 1260
actgggaaga tgtgtagccg aggctggaca ctggcagatg cggatgtggt ttgcagacag 1320
cttggatgtg gatctgcgct tcaaacccag gctaagatct actctaaaac tggggcaaca 1380
aatacgtggc tctttcctgg atcttgtaat ggaaatgaaa ctactttttg gcaatgcaaa 1440
aactggcagt ggggcggcct ttcctgtgat aatttcgaag aagccaaagt tacctgctca 1500
ggccacaggg aacccagact ggttggagga gaaatcccat gctctggtcg tgtggaagtg 1560
aaacacggag acgtgtgggg ctccgtctgt gattttgact tgtctctgga agctgccagt 1620
gtggtgtgca gggaattaca atgtggaaca gtcgtctcta tcctaggggg agcacatttt 1680
ggagaaggaa gtggacagat ctggggtgaa gaattccagt gtagtgggga tgagtcccat 1740
ctttcactat gctcagtggc gcccccgcta gacagaactt gtacccacag cagggatgtc 1800
agcgtagtct gctcacgata catagatatt cgtctggcag gcggcgagtc ctcctgtgag 1860
ggaagagtgg agctcaagac actcggagcc tggggtcccc tctgcagttc tcattgggac 1920
atggaagatg ctcatgtctt atgtcagcag ctgaagtgtg gggttgccca atctattcca 1980
gaaggagcac attttgggaa aggagctggt caggtctgga gtcacatgtt ccactgcact 2040
ggaactgagg aacatatagg agattgcctc atgactgctc tgggtgcgcc gacgtgttcc 2100
gaaggacagg tggcctctgt catctgctca ggaaaccaat cccagacact attgccatgt 2160
agttcattgt ctccagtcca aacaacaagc tctacaattc caaaggagag tgaagttccc 2220
tgcatagcaa gtggccagct tcgcttggta ggtggaggtg gtcgctgcgc tggaagagtg 2280
gaggtctgcc acgagggctc ttggggcacc gtctgtgatg acaattggga tatgactgat 2340
gccaatgtgg tgtgcaagca gctggactgt ggcgtggcaa ttaacgccac tggctctgct 2400
tacttcgggg aaggagcagg agctatctgg ctagacgaag tcatctgcac tgggaaagag 2460
tctcatattt ggcagtgcca ttcacatggc tggggacgcc ataactgcag gcacaaagaa 2520
gatgcaggtg ttatctgctc cgagttcatg tctctgaggc tgaccaacga agcccacaaa 2580
gaaaactgca caggtcgcct tgaagtgttt tacaatggta catggggcag tattggcagt 2640
agcaatatgt ctccaaccac tgtgggggtg gtgtgccgtc agctgggctg tgcagacaac 2700
gggactgtga aacccatacc ttcagacaag acaccatcca ggcccatgtg ggtagatcgt 2760
gtgcagtgtc caaaaggagt tgacactttg tggcagtgcc cctcgtcacc ttggaaacag 2820
agacaggcca gcccctcctc ccaggagtcc tggatcatct gtgacaacaa aataagactc 2880
caggaagggc atacagactg ttctggacgt gtggagatct ggcacaaagg ttcctgggga 2940
acagtgtgtg atgactcctg ggatcttaat gatgctaagg ttgtatgtaa gcagttgggc 3000
tgtggccaag ctgtgaaggc actaaaagaa gcagcatttg gtccaggaac tgggcccata 3060
tggctcaatg aaattaagtg tagagggaat gagtcttccc tgtgggattg tcctgccaaa 3120
ccgtggagtc acagcgactg tgggcacaaa gaagatgctt ccatccagtg cctcccaaaa 3180
atgacttcag aatcacatca tggcacaggt caccccaccc tcacggcact cttggtttgt 3240
ggagccattc tattggtcct cctcattgtc ttcctcctgt ggactctgaa gcgacgacgg 3300
attcagcgac ttacagtttc ctcaagagga gaggtcttga tacatcaagt tcagtaccaa 3360
gagatggatt caaaggcgga tgatctggac ttgctgaaat cctcgggggt cattcagagg 3420
cacactgaga aggaaaatga taatttataa tccactgagg ttggagttta agaagccttg 3480
acaggacagc cagctaaatg gaacaagagc ccaggcaacg cacggatgac cacagctgca 3540
tcttcatgca gtcctttgtt tcctggaact ctgctgaacc tgcaaaaacc atatttgtga 3600
atgtgaccac ttaatagaga tgggagactt ttgagggaat taaacaatat tgctattggt 3660
ttgcttgttc gcaatagggt ctcattatgt atagccctgg agatggcgat agagagcagg 3720
caagcctaga attcacagag atctgcttgt ctctgcttcc caaatgctgg gatcaaatat 3780
gtggaccacc acatgtggtt taacaattgt gtcttgattt tataaatttc tggttggttt 3840
ttctgacgtt tttagggttt cgtgaatata aaataatgtc ttttcggttg gcatgctaat 3900
ttttaaatat tatacacttc cttgtagtga gtttaagaat aatttcttat aaccaagtca 3960
attcattttc actttgaatt atttaataaa ggaatatggt cattgtgacc acacacacag 4020
cagttgtgac cgcctgtatg aggccttcaa aaaatatttt aaaaatagag ggctggagaa 4080
atggctcaga ggtcctgagt tcaattccta gcaaccacat ggtggctcac aaccatctgt 4140
aatgggaatc cgatgccctc ttccagtgta tctgaagata gtgacactgt cctcattaac 4200
ataaaataaa taaataaatc tttaaaaaaa gaaaagacaa tagaggaggg gaggggcgtg 4260
agcgtaggag tgaggactga ttgagaagaa ggttggagga agtgggggga ggtgaaaagc 4320
tagttgggaa cttatgtgat cacagtgcat catgtccaaa tatgacatgt ccaaaaatgt 4380
tattaataaa gaaacggaaa tcaaa 4405
<210> SEQ ID NO 4
<211> LENGTH: 1121
<212> TYPE: PRT
<213> ORGANISM: Mus musculus
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/AAK16065
<309> DATABASE ENTRY DATE: 2001-05-10
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1121)
<400> SEQUENCE: 4
Met Gly Gly His Arg Met Val Leu Leu Gly Gly Ala Gly Ser Pro Gly
1 5 10 15
Cys Lys Arg Phe Val His Leu Gly Phe Phe Val Val Ala Val Ser Ser
20 25 30
Leu Leu Ser Ala Ser Ala Val Thr Asn Ala Pro Gly Glu Met Lys Lys
35 40 45
Glu Leu Arg Leu Ala Gly Gly Glu Asn Asn Cys Ser Gly Arg Val Glu
50 55 60
Leu Lys Ile His Asp Lys Trp Gly Thr Val Cys Ser Asn Gly Trp Ser
65 70 75 80
Met Asn Glu Val Ser Val Val Cys Gln Gln Leu Gly Cys Pro Thr Ser
85 90 95
Ile Lys Ala Leu Gly Trp Ala Asn Ser Ser Ala Gly Ser Gly Tyr Ile
100 105 110
Trp Met Asp Lys Val Ser Cys Thr Gly Asn Glu Ser Ala Leu Trp Asp
115 120 125
Cys Lys His Asp Gly Trp Gly Lys His Asn Cys Thr His Glu Lys Asp
130 135 140
Ala Gly Val Thr Cys Ser Asp Gly Ser Asn Leu Glu Met Arg Leu Val
145 150 155 160
Asn Ser Ala Gly His Arg Cys Leu Gly Arg Val Glu Ile Lys Phe Gln
165 170 175
Gly Lys Trp Gly Thr Val Cys Asp Asp Asn Phe Ser Lys Asp His Ala
180 185 190
Ser Val Ile Cys Lys Gln Leu Gly Cys Gly Ser Ala Ile Ser Phe Ser
195 200 205
Gly Ser Ala Lys Leu Gly Ala Gly Ser Gly Pro Ile Trp Leu Asp Asp
210 215 220
Leu Ala Cys Asn Gly Asn Glu Ser Ala Leu Trp Asp Cys Lys His Arg
225 230 235 240
Gly Trp Gly Lys His Asn Cys Asp His Ala Glu Asp Val Gly Val Ile
245 250 255
Cys Leu Glu Gly Ala Asp Leu Ser Leu Arg Leu Val Asp Gly Val Ser
260 265 270
Arg Cys Ser Gly Arg Leu Glu Val Arg Phe Gln Gly Glu Trp Gly Thr
275 280 285
Val Cys Asp Asp Asn Trp Asp Leu Arg Asp Ala Ser Val Val Cys Lys
290 295 300
Gln Leu Gly Cys Pro Thr Ala Ile Ser Ala Ile Gly Arg Val Asn Ala
305 310 315 320
Ser Glu Gly Ser Gly Gln Ile Trp Leu Asp Asn Ile Ser Cys Glu Gly
325 330 335
His Glu Ala Thr Leu Trp Glu Cys Lys His Gln Glu Trp Gly Lys His
340 345 350
Tyr Cys His His Arg Glu Asp Ala Gly Val Thr Cys Ser Asp Gly Ala
355 360 365
Asp Leu Glu Leu Arg Leu Val Gly Gly Gly Ser Arg Cys Ala Gly Ile
370 375 380
Val Glu Val Glu Ile Gln Lys Leu Thr Gly Lys Met Cys Ser Arg Gly
385 390 395 400
Trp Thr Leu Ala Asp Ala Asp Val Val Cys Arg Gln Leu Gly Cys Gly
405 410 415
Ser Ala Leu Gln Thr Gln Ala Lys Ile Tyr Ser Lys Thr Gly Ala Thr
420 425 430
Asn Thr Trp Leu Phe Pro Gly Ser Cys Asn Gly Asn Glu Thr Thr Phe
435 440 445
Trp Gln Cys Lys Asn Trp Gln Trp Gly Gly Leu Ser Cys Asp Asn Phe
450 455 460
Glu Glu Ala Lys Val Thr Cys Ser Gly His Arg Glu Pro Arg Leu Val
465 470 475 480
Gly Gly Glu Ile Pro Cys Ser Gly Arg Val Glu Val Lys His Gly Asp
485 490 495
Val Trp Gly Ser Val Cys Asp Phe Asp Leu Ser Leu Glu Ala Ala Ser
500 505 510
Val Val Cys Arg Glu Leu Gln Cys Gly Thr Val Val Ser Ile Leu Gly
515 520 525
Gly Ala His Phe Gly Glu Gly Ser Gly Gln Ile Trp Gly Glu Glu Phe
530 535 540
Gln Cys Ser Gly Asp Glu Ser His Leu Ser Leu Cys Ser Val Ala Pro
545 550 555 560
Pro Leu Asp Arg Thr Cys Thr His Ser Arg Asp Val Ser Val Val Cys
565 570 575
Ser Arg Tyr Ile Asp Ile Arg Leu Ala Gly Gly Glu Ser Ser Cys Glu
580 585 590
Gly Arg Val Glu Leu Lys Thr Leu Gly Ala Trp Gly Pro Leu Cys Ser
595 600 605
Ser His Trp Asp Met Glu Asp Ala His Val Leu Cys Gln Gln Leu Lys
610 615 620
Cys Gly Val Ala Gln Ser Ile Pro Glu Gly Ala His Phe Gly Lys Gly
625 630 635 640
Ala Gly Gln Val Trp Ser His Met Phe His Cys Thr Gly Thr Glu Glu
645 650 655
His Ile Gly Asp Cys Leu Met Thr Ala Leu Gly Ala Pro Thr Cys Ser
660 665 670
Glu Gly Gln Val Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln Thr
675 680 685
Leu Leu Pro Cys Ser Ser Leu Ser Pro Val Gln Thr Thr Ser Ser Thr
690 695 700
Ile Pro Lys Glu Ser Glu Val Pro Cys Ile Ala Ser Gly Gln Leu Arg
705 710 715 720
Leu Val Gly Gly Gly Gly Arg Cys Ala Gly Arg Val Glu Val Tyr His
725 730 735
Glu Gly Ser Trp Gly Thr Val Cys Asp Asp Asn Trp Asp Met Thr Asp
740 745 750
Ala Asn Val Val Cys Lys Gln Leu Asp Cys Gly Val Ala Ile Asn Ala
755 760 765
Thr Gly Ser Ala Tyr Phe Gly Glu Gly Ala Gly Ala Ile Trp Leu Asp
770 775 780
Glu Val Ile Cys Thr Gly Lys Glu Ser His Ile Trp Gln Cys His Ser
785 790 795 800
His Gly Trp Gly Arg His Asn Cys Arg His Lys Glu Asp Ala Gly Val
805 810 815
Ile Cys Ser Glu Phe Met Ser Leu Arg Leu Thr Asn Glu Ala His Lys
820 825 830
Glu Ser Cys Thr Gly Arg Leu Glu Val Phe Tyr Asn Gly Thr Trp Gly
835 840 845
Ser Ile Gly Ser Ser Asn Met Ser Pro Thr Thr Val Gly Val Val Cys
850 855 860
Arg Gln Leu Gly Cys Ala Asp Asn Gly Thr Val Lys Pro Ile Pro Ser
865 870 875 880
Asp Lys Thr Pro Ser Arg Pro Met Trp Val Asp Arg Val Gln Cys Pro
885 890 895
Lys Gly Val Asp Thr Leu Trp Gln Cys Pro Ser Ser Pro Trp Lys Gln
900 905 910
Arg Gln Ala Ser Pro Ser Ser Gln Glu Ser Trp Ile Ile Cys Asp Asn
915 920 925
Lys Ile Arg Leu Gln Glu Gly His Thr Asp Cys Ser Gly Arg Val Glu
930 935 940
Ile Trp His Lys Gly Phe Trp Gly Thr Val Cys Asp Asp Ser Trp Asp
945 950 955 960
Leu Asn Asp Ala Lys Val Val Cys Lys Gln Leu Gly Cys Gly Gln Ala
965 970 975
Val Lys Ala Leu Lys Glu Ala Ala Phe Gly Pro Gly Thr Gly Pro Ile
980 985 990
Trp Leu Asn Glu Ile Lys Cys Arg Gly Asn Glu Ser Ser Leu Trp Asp
995 1000 1005
Cys Pro Ala Lys Pro Trp Ser His Ser Asp Cys Gly His Lys Glu
1010 1015 1020
Asp Ala Ser Ile Gln Cys Leu Pro Lys Met Thr Ser Glu Ser His
1025 1030 1035
His Gly Thr Gly His Pro Thr Leu Thr Ala Leu Leu Val Cys Gly
1040 1045 1050
Ala Ile Leu Leu Val Leu Leu Ile Val Phe Leu Leu Trp Thr Leu
1055 1060 1065
Lys Arg Arg Gln Ile Gln Arg Leu Thr Val Ser Ser Arg Gly Glu
1070 1075 1080
Val Leu Ile His Gln Val Gln Tyr Gln Glu Met Asp Ser Lys Ala
1085 1090 1095
Asp Asp Leu Asp Leu Leu Lys Ser Ser Gly Val Ile Gln Arg His
1100 1105 1110
Thr Glu Lys Glu Asn Asp Asn Leu
1115 1120
<210> SEQ ID NO 5
<211> LENGTH: 3780
<212> TYPE: DNA
<213> ORGANISM: homo sapiens
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/BC051281
<309> DATABASE ENTRY DATE: 2006-07-15
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(3780)
<400> SEQUENCE: 5
aacatttcta gggaataata caagaagatt taggaatcat tgaagttata aatctttgga 60
atgagcaaac tcagaatggt gctacttgaa gactctggat ctgctgactt cagaagacat 120
tttgtcaact tgagtccctt caccattact gtggtcttac ttctcagtgc ctgttttgtc 180
accagttctc ttggaggaac agacaaggag ctgaggctag tggatggtga aaacaagtgt 240
agcgggagag tggaagtgaa agtccaggag gagtggggaa cggtgtgtaa taatggctgg 300
agcatggaag cggtctctgt gatttgtaac cagctgggat gtccaactgc tatcaaagcc 360
cctggatggg ctaattccag tgcaggttct ggacgcattt ggatggatca tgtttcttgt 420
cgtgggaatg agtcagctct ttgggattgc aaacatgatg gatggggaaa gcatagtaac 480
tgtactcacc aacaagatgc tggagtgacc tgctcagatg gatccaattt ggaaatgagg 540
ctgacgcgtg gagggaatat gtgttctgga agaatagaga tcaaattcca aggacggtgg 600
ggaacagtgt gtgatgataa cttcaacata gatcatgcat ctgtcatttg tagacaactt 660
gaatgtggaa gtgctgtcag tttctctggt tcatctaatt ttggagaagg ctctggacca 720
atctggtttg atgatcttat atgcaacgga aatgagtcag ctctctggaa ctgcaaacat 780
caaggatggg gaaagcataa ctgtgatcat gctgaggatg ctggagtgat ttgctcaaag 840
ggagcagatc tgagcctgag actggtagat ggagtcactg aatgttcagg aagattagaa 900
gtgagattcc aaggagaatg ggggacaata tgtgatgacg gctgggacag ttacgatgct 960
gctgtggcat gcaagcaact gggatgtcca actgccgtca cagccattgg tcgagttaac 1020
gccagtaagg gatttggaca catctggctt gacagcgttt cttgccaggg acatgaacct 1080
gctgtctggc aatgtaaaca ccatgaatgg ggaaagcatt attgcaatca caatgaagat 1140
gctggcgtga catgttctga tggatcagat ctggagctaa gacttagagg tggaggcagc 1200
cgctgtgctg ggacagttga ggtggagatt cagagactgt tagggaaggt gtgtgacaga 1260
ggctggggac tgaaagaagc tgatgtggtt tgcaggcagc tgggatgtgg atctgcactc 1320
aaaacatctt atcaagtgta ctccaaaatc caggcaacaa acacatggct gtttctaagt 1380
agctgtaacg gaaatgaaac ttctctttgg gactgcaaga actggcaatg gggtggactt 1440
acctgtgatc actatgaaga agccaaaatt acctgctcag cccacaggga acccagactg 1500
gttggagggg acattccctg ttctggacgt gttgaagtga agcatggtga cacgtggggc 1560
tccatctgtg attcggactt ctctctggaa gctgccagcg ttctatgcag ggaattacag 1620
tgtggcacag ttgtctctat cctgggggga gctcactttg gagagggaaa tggacagatc 1680
tgggctgaag aattccagtg tgagggacat gagtcccatc tttcactctg cccagtagca 1740
ccccgcccag aaggaacttg tagccacagc agggatgttg gagtagtctg ctcaagatac 1800
acagaaattc gcttggtgaa tggcaagacc ccgtgtgagg gcagagtgga gctcaaaacg 1860
cttggtgcct ggggatccct ctgtaactct cactgggaca tagaagatgc ccatgttctt 1920
tgccagcagc ttaaatgtgg agttgccctt tctaccccag gaggagcacg ttttggaaaa 1980
ggaaatggtc agatctggag gcatatgttt cactgcactg ggactgagca gcacatggga 2040
gattgtcctg taactgctct aggtgcttca ttatgtcctt cagagcaagt ggcctctgta 2100
atctgctcag gaaaccagtc ccaaacactg tcctcgtgca attcatcgtc tttgggccca 2160
acaaggccta ccattccaga agaaagtgct gtggcctgca tagagagtgg tcaacttcgc 2220
ctggtaaatg gaggaggtcg ctgtgctggg agagtagaga tctatcatga gggctcctgg 2280
ggcaccatct gtgatgacag ctgggacctg agtgatgccc acgtggtttg cagacagctg 2340
ggctgtggag aggccattaa tgccactggt tctgctcatt ttggggaagg aacagggccc 2400
atctggctgg atgagatgaa atgcaatgga aaagaatccc gcatttggca gtgccattca 2460
cacggctggg ggcagcaaaa ttgcaggcac aaggaggatg cgggagttat ctgctcagaa 2520
ttcatgtctc tgagactgac cagtgaagcc agcagagagg cctgtgcagg gcgtctggaa 2580
gttttttaca atggagcttg gggcactgtt ggcaagagta gcatgtctga aaccactgtg 2640
ggtgtggtgt gcaggcagct gggctgtgca gacaaaggga aaatcaaccc tgcatcttta 2700
gacaaggcca tgtccattcc catgtgggtg gacaatgttc agtgtccaaa aggacctgac 2760
acgctgtggc agtgcccatc atctccatgg gagaagagac tggccagccc ctcggaggag 2820
acctggatca catgtgacaa caagataaga cttcaggaag gacccacttc ctgttctgga 2880
cgtgtggaga tctggcatgg aggttcctgg gggacagtgt gtgatgactc ttgggacttg 2940
gacgatgctc aggtggtgtg tcaacaactt ggctgtggtc cagctttgaa agcattcaaa 3000
gaagcagagt ttggtcaggg gactggaccg atatggctca atgaagtgaa gtgcaaaggg 3060
aatgagtctt ccttgtggga ttgtcctgcc agacgctggg gccatagtga gtgtgggcac 3120
aaggaagacg ctgcagtgaa ttgcacagat atttcagtgc agaaaacccc acaaaaagcc 3180
acaacaggtc gctcatcccg tcagtcatcc tttattgcag tcgggatcct tggggttgtt 3240
ctgttggcca ttttcgtcgc attattcttc ttgactaaaa agcgaagaca gagacagcgg 3300
cttgcagttt cctcaagagg agagaactta gtccaccaaa ttcaataccg ggagatgaat 3360
tcttgcctga atgcagatga tctggaccta atgaattcct cagaaaattc ccatgagtca 3420
gctgatttca gtgctgctga actaatttct gtgtctaaat ttcttcctat ttctggaatg 3480
gaaaaggagg ccattctgag ccacactgaa aaggaaaatg ggaatttata acccagtgag 3540
ttcagccttt aagatacctt gatgaagacc tggactattg aatggagcag aaattcacct 3600
ctctcactga ctattacagt tgcattttta tggagttctt cttctcctag gattcctaag 3660
actgctgctg aatttataaa aattaagttt gtgaatgtga ctacttagtg gtgtatatga 3720
gactttcaag ggaattaaat aaataaataa gaatgttatt gaaaaaaaaa aaaaaaaaaa 3780
<210> SEQ ID NO 6
<211> LENGTH: 1156
<212> TYPE: PRT
<213> ORGANISM: HOMO SAPIENS
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/AAH51281
<309> DATABASE ENTRY DATE: 2006-07-15
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1156)
<400> SEQUENCE: 6
Met Ser Lys Leu Arg Met Val Leu Leu Glu Asp Ser Gly Ser Ala Asp
1 5 10 15
Phe Arg Arg His Phe Val Asn Leu Ser Pro Phe Thr Ile Thr Val Val
20 25 30
Leu Leu Leu Ser Ala Cys Phe Val Thr Ser Ser Leu Gly Gly Thr Asp
35 40 45
Lys Glu Leu Arg Leu Val Asp Gly Glu Asn Lys Cys Ser Gly Arg Val
50 55 60
Glu Val Lys Val Gln Glu Glu Trp Gly Thr Val Cys Asn Asn Gly Trp
65 70 75 80
Ser Met Glu Ala Val Ser Val Ile Cys Asn Gln Leu Gly Cys Pro Thr
85 90 95
Ala Ile Lys Ala Pro Gly Trp Ala Asn Ser Ser Ala Gly Ser Gly Arg
100 105 110
Ile Trp Met Asp His Val Ser Cys Arg Gly Asn Glu Ser Ala Leu Trp
115 120 125
Asp Cys Lys His Asp Gly Trp Gly Lys His Ser Asn Cys Thr His Gln
130 135 140
Gln Asp Ala Gly Val Thr Cys Ser Asp Gly Ser Asn Leu Glu Met Arg
145 150 155 160
Leu Thr Arg Gly Gly Asn Met Cys Ser Gly Arg Ile Glu Ile Lys Phe
165 170 175
Gln Gly Arg Trp Gly Thr Val Cys Asp Asp Asn Phe Asn Ile Asp His
180 185 190
Ala Ser Val Ile Cys Arg Gln Leu Glu Cys Gly Ser Ala Val Ser Phe
195 200 205
Ser Gly Ser Ser Asn Phe Gly Glu Gly Ser Gly Pro Ile Trp Phe Asp
210 215 220
Asp Leu Ile Cys Asn Gly Asn Glu Ser Ala Leu Trp Asn Cys Lys His
225 230 235 240
Gln Gly Trp Gly Lys His Asn Cys Asp His Ala Glu Asp Ala Gly Val
245 250 255
Ile Cys Ser Lys Gly Ala Asp Leu Ser Leu Arg Leu Val Asp Gly Val
260 265 270
Thr Glu Cys Ser Gly Arg Leu Glu Val Arg Phe Gln Gly Glu Trp Gly
275 280 285
Thr Ile Cys Asp Asp Gly Trp Asp Ser Tyr Asp Ala Ala Val Ala Cys
290 295 300
Lys Gln Leu Gly Cys Pro Thr Ala Val Thr Ala Ile Gly Arg Val Asn
305 310 315 320
Ala Ser Lys Gly Phe Gly His Ile Trp Leu Asp Ser Val Ser Cys Gln
325 330 335
Gly His Glu Pro Ala Val Trp Gln Cys Lys His His Glu Trp Gly Lys
340 345 350
His Tyr Cys Asn His Asn Glu Asp Ala Gly Val Thr Cys Ser Asp Gly
355 360 365
Ser Asp Leu Glu Leu Arg Leu Arg Gly Gly Gly Ser Arg Cys Ala Gly
370 375 380
Thr Val Glu Val Glu Ile Gln Arg Leu Leu Gly Lys Val Cys Asp Arg
385 390 395 400
Gly Trp Gly Leu Lys Glu Ala Asp Val Val Cys Arg Gln Leu Gly Cys
405 410 415
Gly Ser Ala Leu Lys Thr Ser Tyr Gln Val Tyr Ser Lys Ile Gln Ala
420 425 430
Thr Asn Thr Trp Leu Phe Leu Ser Ser Cys Asn Gly Asn Glu Thr Ser
435 440 445
Leu Trp Asp Cys Lys Asn Trp Gln Trp Gly Gly Leu Thr Cys Asp His
450 455 460
Tyr Glu Glu Ala Lys Ile Thr Cys Ser Ala His Arg Glu Pro Arg Leu
465 470 475 480
Val Gly Gly Asp Ile Pro Cys Ser Gly Arg Val Glu Val Lys His Gly
485 490 495
Asp Thr Trp Gly Ser Ile Cys Asp Ser Asp Phe Ser Leu Glu Ala Ala
500 505 510
Ser Val Leu Cys Arg Glu Leu Gln Cys Gly Thr Val Val Ser Ile Leu
515 520 525
Gly Gly Ala His Phe Gly Glu Gly Asn Gly Gln Ile Trp Ala Glu Glu
530 535 540
Phe Gln Cys Glu Gly His Glu Ser His Leu Ser Leu Cys Pro Val Ala
545 550 555 560
Pro Arg Pro Glu Gly Thr Cys Ser His Ser Arg Asp Val Gly Val Val
565 570 575
Cys Ser Arg Tyr Thr Glu Ile Arg Leu Val Asn Gly Lys Thr Pro Cys
580 585 590
Glu Gly Arg Val Glu Leu Lys Thr Leu Gly Ala Trp Gly Ser Leu Cys
595 600 605
Asn Ser His Trp Asp Ile Glu Asp Ala His Val Leu Cys Gln Gln Leu
610 615 620
Lys Cys Gly Val Ala Leu Ser Thr Pro Gly Gly Ala Arg Phe Gly Lys
625 630 635 640
Gly Asn Gly Gln Ile Trp Arg His Met Phe His Cys Thr Gly Thr Glu
645 650 655
Gln His Met Gly Asp Cys Pro Val Thr Ala Leu Gly Ala Ser Leu Cys
660 665 670
Pro Ser Glu Gln Val Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln
675 680 685
Thr Leu Ser Ser Cys Asn Ser Ser Ser Leu Gly Pro Thr Arg Pro Thr
690 695 700
Ile Pro Glu Glu Ser Ala Val Ala Cys Ile Glu Ser Gly Gln Leu Arg
705 710 715 720
Leu Val Asn Gly Gly Gly Arg Cys Ala Gly Arg Val Glu Ile Tyr His
725 730 735
Glu Gly Ser Trp Gly Thr Ile Cys Asp Asp Ser Trp Asp Leu Ser Asp
740 745 750
Ala His Val Val Cys Arg Gln Leu Gly Cys Gly Glu Ala Ile Asn Ala
755 760 765
Thr Gly Ser Ala His Phe Gly Glu Gly Thr Gly Pro Ile Trp Leu Asp
770 775 780
Glu Met Lys Cys Asn Gly Lys Glu Ser Arg Ile Trp Gln Cys His Ser
785 790 795 800
His Gly Trp Gly Gln Gln Asn Cys Arg His Lys Glu Asp Ala Gly Val
805 810 815
Ile Cys Ser Glu Phe Met Ser Leu Arg Leu Thr Ser Glu Ala Ser Arg
820 825 830
Glu Ala Cys Ala Gly Arg Leu Glu Val Phe Tyr Asn Gly Ala Trp Gly
835 840 845
Thr Val Gly Lys Ser Ser Met Ser Glu Thr Thr Val Gly Val Val Cys
850 855 860
Arg Gln Leu Gly Cys Ala Asp Lys Gly Lys Ile Asn Pro Ala Ser Leu
865 870 875 880
Asp Lys Ala Met Ser Ile Pro Met Trp Val Asp Asn Val Gln Cys Pro
885 890 895
Lys Gly Pro Asp Thr Leu Trp Gln Cys Pro Ser Ser Pro Trp Glu Lys
900 905 910
Arg Leu Ala Ser Pro Ser Glu Glu Thr Trp Ile Thr Cys Asp Asn Lys
915 920 925
Ile Arg Leu Gln Glu Gly Pro Thr Ser Cys Ser Gly Arg Val Glu Ile
930 935 940
Trp His Gly Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu
945 950 955 960
Asp Asp Ala Gln Val Val Cys Gln Gln Leu Gly Cys Gly Pro Ala Leu
965 970 975
Lys Ala Phe Lys Glu Ala Glu Phe Gly Gln Gly Thr Gly Pro Ile Trp
980 985 990
Leu Asn Glu Val Lys Cys Lys Gly Asn Glu Ser Ser Leu Trp Asp Cys
995 1000 1005
Pro Ala Arg Arg Trp Gly His Ser Glu Cys Gly His Lys Glu Asp
1010 1015 1020
Ala Ala Val Asn Cys Thr Asp Ile Ser Val Gln Lys Thr Pro Gln
1025 1030 1035
Lys Ala Thr Thr Gly Arg Ser Ser Arg Gln Ser Ser Phe Ile Ala
1040 1045 1050
Val Gly Ile Leu Gly Val Val Leu Leu Ala Ile Phe Val Ala Leu
1055 1060 1065
Phe Phe Leu Thr Lys Lys Arg Arg Gln Arg Gln Arg Leu Ala Val
1070 1075 1080
Ser Ser Arg Gly Glu Asn Leu Val His Gln Ile Gln Tyr Arg Glu
1085 1090 1095
Met Asn Ser Cys Leu Asn Ala Asp Asp Leu Asp Leu Met Asn Ser
1100 1105 1110
Ser Glu Asn Ser His Glu Ser Ala Asp Phe Ser Ala Ala Glu Leu
1115 1120 1125
Ile Ser Val Ser Lys Phe Leu Pro Ile Ser Gly Met Glu Lys Glu
1130 1135 1140
Ala Ile Leu Ser His Thr Glu Lys Glu Asn Gly Asn Leu
1145 1150 1155
<210> SEQ ID NO 7
<211> LENGTH: 4950
<212> TYPE: DNA
<213> ORGANISM: homo sapiens
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/Z22970
<309> DATABASE ENTRY DATE: 2005-04-18
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(4950)
<400> SEQUENCE: 7
gaattcttag ttgttttctt tagaagaaca tttctaggga ataatacaag aagatttagg 60
aatcattgaa gttataaatc tttggaatga gcaaactcag aatggtgcta cttgaagact 120
ctggatctgc tgacttcaga agacattttg tcaacctgag tcccttcacc attactgtgg 180
tcttacttct cagtgcctgt tttgtcacca gttctcttgg aggaacagac aaggagctga 240
ggctagtgga tggtgaaaac aagtgtagcg ggagagtgga agtgaaagtc caggaggagt 300
ggggaacggt gtgtaataat ggctggagca tggaagcggt ctctgtgatt tgtaaccagc 360
tgggatgtcc aactgctatc aaagcccctg gatgggctaa ttccagtgca ggttctggac 420
gcatttggat ggatcatgtt tcttgtcgtg ggaatgagtc agctctttgg gattgcaaac 480
atgatggatg gggaaagcat agtaactgta ctcaccaaca agatgctgga gtgacctgct 540
cagatggatc caatttggaa atgaggctga cgcgtggagg gaatatgtgt tctggaagaa 600
tagagatcaa attccaagga cggtggggaa cagtgtgtga tgataacttc aacatagatc 660
atgcatctgt catttgtaga caacttgaat gtggaagtgc tgtcagtttc tctggttcat 720
ctaattttgg agaaggctct ggaccaatct ggtttgatga tcttatatgc aacggaaatg 780
agtcagctct ctggaactgc aaacatcaag gatggggaaa gcataactgt gatcatgctg 840
aggatgctgg agtgatttgc tcaaagggag cagatctgag cctgagactg gtagatggag 900
tcactgaatg ttcaggaaga ttagaagtga gattccaagg agaatggggg acaatatgtg 960
atgacggctg ggacagttac gatgctgctg tggcatgcaa gcaactggga tgtccaactg 1020
ccgtcacagc cattggtcga gttaacgcca gtaagggatt tggacacatc tggcttgaca 1080
gcgtttcttg ccagggacat gaacctgctg tctggcaatg taaacaccat gaatggggaa 1140
agcattattg caatcacaat gaagatgctg gcgtgacatg ttctgatgga tcagatctgg 1200
agctaagact tagaggtgga ggcagccgct gtgctgggac agttgaggtg gagattcaga 1260
gactgttagg gaaggtgtgt gacagaggct ggggactgaa agaagctgat gtggtttgca 1320
ggcagctggg atgtggatct gcactcaaaa catcttatca agtgtactcc aaaatccagg 1380
caacaaacac atggctgttt ctaagtagct gtaacggaaa tgaaacttct ctttgggact 1440
gcaagaactg gcaatggggt ggacttacct gtgatcacta tgaagaagcc aaaattacct 1500
gctcagccca cagggaaccc agactggttg gaggggacat tccctgttct ggacgtgttg 1560
aagtgaagca tggtgacacg tggggctcca tctgtgattc ggacttctct ctggaagctg 1620
ccagcgttct atgcagggaa ttacagtgtg gcacagttgt ctctatcctg gggggagctc 1680
actttggaga gggaaatgga cagatctggg ctgaagaatt ccagtgtgag ggacatgagt 1740
cccatctttc actctgccca gtagcacccc gcccagaagg aacttgtagc cacagcaggg 1800
atgttggagt agtctgctca agatacacag aaattcgctt ggtgaatggc aagaccccgt 1860
gtgagggcag agtggagctc aaaacgcttg gtgcctgggg atccctctgt aactctcact 1920
gggacataga agatgcccat gttctttgcc agcagcttaa atgtggagtt gccctttcta 1980
ccccaggagg agcacgtttt ggaaaaggaa atggtcagat ctggaggcat atgtttcact 2040
gcactgggac tgagcagcac atgggagatt gtcctgtaac tgctctaggt gcttcattat 2100
gtccttcaga gcaagtggcc tctgtaatct gctcaggaaa ccagtcccaa acactgtcct 2160
cgtgcaattc atcgtctttg ggcccaacaa ggcctaccat tccagaagaa agtgctgtgg 2220
cctgcataga gagtggtcaa cttcgcctgg taaatggagg aggtcgctgt gctgggagag 2280
tagagatcta tcatgagggc tcctggggca ccatctgtga tgacagctgg gacctgagtg 2340
atgcccacgt ggtttgcaga cagctgggct gtggagaggc cattaatgcc actggttctg 2400
ctcattttgg ggaaggaaca gggcccatct ggctggatga gatgaaatgc aatggaaaag 2460
aatcccgcat ttggcagtgc cattcacacg gctgggggca gcaaaattgc aggcacaagg 2520
aggatgcggg agttatctgc tcagaattca tgtctctgag actgaccagt gaagccagca 2580
gagaggcctg tgcagggcgt ctggaagttt tttacaatgg agcttggggc actgttggca 2640
agagtagcat gtctgaaacc actgtgggtg tggtgtgcag gcagctgggc tgtgcagaca 2700
aagggaaaat caaccctgca tctttagaca aggccatgtc cattcccatg tgggtggaca 2760
atgttcagtg tccaaaagga cctgacacgc tgtggcagtg cccatcatct ccatgggaga 2820
agagactggc cagcccctcg gaggagacct ggatcacatg tgacaacaag ataagacttc 2880
aggaaggacc cacttcctgt tctggacgtg tggagatctg gcatggaggt tcctggggga 2940
cagtgtgtga tgactcttgg gacttggacg atgctcaggt ggtgtgtcaa caacttggct 3000
gtggtccagc tttgaaagca ttcaaagaag cagagtttgg tcaggggact ggaccgatat 3060
ggctcaatga agtgaagtgc aaagggaatg agtcttcctt gtgggattgt cctgccagac 3120
gctggggcca tagtgagtgt gggcacaagg aagacgctgc agtgaattgc acagatattt 3180
cagtgcagaa aaccccacaa aaagccacaa caggtcgctc atcccgtcag tcatccttta 3240
ttgcagtcgg gatccttggg gttgttctgt tggccatttt cgtcgcatta ttcttcttga 3300
ctaaaaagcg aagacagaga cagcggcttg cagtttcctc aagaggagag aacttagtcc 3360
accaaattca ataccgggag atgaattctt gcctgaatgc agatgatctg gacctaatga 3420
attcctcagg tctgtgggtt cttggagggt ctattgccca ggggttcaga tcagtggctg 3480
cagttgaggc acagacattc tactttgata aacagttaaa aaagtctaaa aatgtaatag 3540
gaagcttaga tgcatataat ggacaagaat gactgaaaat tattcttgga gaatatcaaa 3600
attgcaatca tagggaggcc tttagcttaa gaggcctgtg attattcctg atagaggtat 3660
ggaaagaacc atgcagagga atattatgac ttggacctca ttttattaaa acagaaatta 3720
atcttacaaa agattgtcat aagtgacagt ttaacttttt tctttaaatt ttgttgtgta 3780
tatttaaggt atacaacatg attttatggg atgtatatag atagtaaaaa gcttactaaa 3840
gcaaagcaaa tgaacacacc catcatctga catagttacc cttttttgtg ttgttcttgt 3900
ggcaagagca gctaaaacct actcacttag catgaatcct acatacagca caatgttatt 3960
acctataatc ctcatgttgt acattagacc tctagactgg ttcattctac gtatctgcta 4020
ctttgtatcc tctgacctac atacgtcttt cacagtttct tccattccca tttcctgtca 4080
ttttttttct ctagcttgat atttattata tttttcccta aaagtctaaa accttaaact 4140
ttcaatatct ttattgcatg agaagccata caaatccaca gaactagcct tatttctcat 4200
cacatcatgc tgttttatcc ttgaacttct atttagcacc agtgcactaa ttctgcatct 4260
gggcaggatg actttactgg gttggaagaa atatcccaaa acccattgtc tttactccat 4320
gaagggtccc tgaccttctg agaggggcct gcctcacttc ttccatccaa agaattatgc 4380
atctgctact gtgtcaggga acatatttaa ggaacatgta ctgttactgt gtcaggaaac 4440
atatttaaga aataggaaag actttctctg ccccttaaat cacacatgct tttcttccta 4500
gttatgggtg gtgtttttag ttgctcaaag agcctcacag ttacgtgaga agaggtctgg 4560
tttatttccc agtaattatt ttcttccttt cagaaaattc ccatgagtca gctgatttca 4620
gtgctgctga actaatttct gtgtctaaat ttcttcctat ttctggaatg gaaaaggagg 4680
ccattctgag ccacactgaa aaggaaaatg ggaatttata acccagtgag ttcagccttt 4740
aagatacctt gatgaagacc tggactattg aatggagcag aaattcacct ctctcactga 4800
ctattacagt tgcattttta tggagttctt cttctcctag gattcctaag actgctgctg 4860
aatttataaa aattaagttt gtgaatgtga ctacttagtg gtgtatatga gactttcaag 4920
ggaattaaat aaataaataa gaatgttaaa 4950
<210> SEQ ID NO 8
<211> LENGTH: 1156
<212> TYPE: PRT
<213> ORGANISM: HOMO SAPIENS
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/CAA80543
<309> DATABASE ENTRY DATE: 2005-04-18
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1156)
<400> SEQUENCE: 8
Met Val Leu Leu Glu Asp Ser Gly Ser Ala Asp Phe Arg Arg His Phe
1 5 10 15
Val Asn Leu Ser Pro Phe Thr Ile Thr Val Val Leu Leu Leu Ser Ala
20 25 30
Cys Phe Val Thr Ser Ser Leu Gly Gly Thr Asp Lys Glu Leu Arg Leu
35 40 45
Val Asp Gly Glu Asn Lys Cys Ser Gly Arg Val Glu Val Lys Val Gln
50 55 60
Glu Glu Trp Gly Thr Val Cys Asn Asn Gly Trp Ser Met Glu Ala Val
65 70 75 80
Ser Val Ile Cys Asn Gln Leu Gly Cys Pro Thr Ala Ile Lys Ala Pro
85 90 95
Gly Trp Ala Asn Ser Ser Ala Gly Ser Gly Arg Ile Trp Met Asp His
100 105 110
Val Ser Cys Arg Gly Asn Glu Ser Ala Leu Trp Asp Cys Lys His Asp
115 120 125
Gly Trp Gly Lys His Ser Asn Cys Thr His Gln Gln Asp Ala Gly Val
130 135 140
Thr Cys Ser Asp Gly Ser Asn Leu Glu Met Arg Leu Thr Arg Gly Gly
145 150 155 160
Asn Met Cys Ser Gly Arg Ile Glu Ile Lys Phe Gln Gly Arg Trp Gly
165 170 175
Thr Val Cys Asp Asp Asn Phe Asn Ile Asp His Ala Ser Val Ile Cys
180 185 190
Arg Gln Leu Glu Cys Gly Ser Ala Val Ser Phe Ser Gly Ser Ser Asn
195 200 205
Phe Gly Glu Gly Ser Gly Pro Ile Trp Phe Asp Asp Leu Ile Cys Asn
210 215 220
Gly Asn Glu Ser Ala Leu Trp Asn Cys Lys His Gln Gly Trp Gly Lys
225 230 235 240
His Asn Cys Asp His Ala Glu Asp Ala Gly Val Ile Cys Ser Lys Gly
245 250 255
Ala Asp Leu Ser Leu Arg Leu Val Asp Gly Val Thr Glu Cys Ser Gly
260 265 270
Arg Leu Glu Val Arg Phe Gln Gly Glu Trp Gly Thr Ile Cys Asp Asp
275 280 285
Gly Trp Asp Ser Tyr Asp Ala Ala Val Ala Cys Lys Gln Leu Gly Cys
290 295 300
Pro Thr Ala Val Thr Ala Ile Gly Arg Val Asn Ala Ser Lys Gly Phe
305 310 315 320
Gly His Ile Trp Leu Asp Ser Val Ser Cys Gln Gly His Glu Pro Ala
325 330 335
Val Trp Gln Cys Lys His His Glu Trp Gly Lys His Tyr Cys Asn His
340 345 350
Asn Glu Asp Ala Gly Val Thr Cys Ser Asp Gly Ser Asp Leu Glu Leu
355 360 365
Arg Leu Arg Gly Gly Gly Ser Arg Cys Ala Gly Thr Val Glu Val Glu
370 375 380
Ile Gln Arg Leu Leu Gly Lys Val Cys Asp Arg Gly Trp Gly Leu Lys
385 390 395 400
Glu Ala Asp Val Val Cys Arg Gln Leu Gly Cys Gly Ser Ala Leu Lys
405 410 415
Thr Ser Tyr Gln Val Tyr Ser Lys Ile Gln Ala Thr Asn Thr Trp Leu
420 425 430
Phe Leu Ser Ser Cys Asn Gly Asn Glu Thr Ser Leu Trp Asp Cys Lys
435 440 445
Asn Trp Gln Trp Gly Gly Leu Thr Cys Asp His Tyr Glu Glu Ala Lys
450 455 460
Ile Thr Cys Ser Ala His Arg Glu Pro Arg Leu Val Gly Gly Asp Ile
465 470 475 480
Pro Cys Ser Gly Arg Val Glu Val Lys His Gly Asp Thr Trp Gly Ser
485 490 495
Ile Cys Asp Ser Asp Phe Ser Leu Glu Ala Ala Ser Val Leu Cys Arg
500 505 510
Glu Leu Gln Cys Gly Thr Val Val Ser Ile Leu Gly Gly Ala His Phe
515 520 525
Gly Glu Gly Asn Gly Gln Ile Trp Ala Glu Glu Phe Gln Cys Glu Gly
530 535 540
His Glu Ser His Leu Ser Leu Cys Pro Val Ala Pro Arg Pro Glu Gly
545 550 555 560
Thr Cys Ser His Ser Arg Asp Val Gly Val Val Cys Ser Arg Tyr Thr
565 570 575
Glu Ile Arg Leu Val Asn Gly Lys Thr Pro Cys Glu Gly Arg Val Glu
580 585 590
Leu Lys Thr Leu Gly Ala Trp Gly Ser Leu Cys Asn Ser His Trp Asp
595 600 605
Ile Glu Asp Ala His Val Leu Cys Gln Gln Leu Lys Cys Gly Val Ala
610 615 620
Leu Ser Thr Pro Gly Gly Ala Arg Phe Gly Lys Gly Asn Gly Gln Ile
625 630 635 640
Trp Arg His Met Phe His Cys Thr Gly Thr Glu Gln His Met Gly Asp
645 650 655
Cys Pro Val Thr Ala Leu Gly Ala Ser Leu Cys Pro Ser Glu Gln Val
660 665 670
Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln Thr Leu Ser Ser Cys
675 680 685
Asn Ser Ser Ser Leu Gly Pro Thr Arg Pro Thr Ile Pro Glu Glu Ser
690 695 700
Ala Val Ala Cys Ile Glu Ser Gly Gln Leu Arg Leu Val Asn Gly Gly
705 710 715 720
Gly Arg Cys Ala Gly Arg Val Glu Ile Tyr His Glu Gly Ser Trp Gly
725 730 735
Thr Ile Cys Asp Asp Ser Trp Asp Leu Ser Asp Ala His Val Val Cys
740 745 750
Arg Gln Leu Gly Cys Gly Glu Ala Ile Asn Ala Thr Gly Ser Ala His
755 760 765
Phe Gly Glu Gly Thr Gly Pro Ile Trp Leu Asp Glu Met Lys Cys Asn
770 775 780
Gly Lys Glu Ser Arg Ile Trp Gln Cys His Ser His Gly Trp Gly Gln
785 790 795 800
Gln Asn Cys Arg His Lys Glu Asp Ala Gly Val Ile Cys Ser Glu Phe
805 810 815
Met Ser Leu Arg Leu Thr Ser Glu Ala Ser Arg Glu Ala Cys Ala Gly
820 825 830
Arg Leu Glu Val Phe Tyr Asn Gly Ala Trp Gly Thr Val Gly Lys Ser
835 840 845
Ser Met Ser Glu Thr Thr Val Gly Val Val Cys Arg Gln Leu Gly Cys
850 855 860
Ala Asp Lys Gly Lys Ile Asn Pro Ala Ser Leu Asp Lys Ala Met Ser
865 870 875 880
Ile Pro Met Trp Val Asp Asn Val Gln Cys Pro Lys Gly Pro Asp Thr
885 890 895
Leu Trp Gln Cys Pro Ser Ser Pro Trp Glu Lys Arg Leu Ala Ser Pro
900 905 910
Ser Glu Glu Thr Trp Ile Thr Cys Asp Asn Lys Ile Arg Leu Gln Glu
915 920 925
Gly Pro Thr Ser Cys Ser Gly Arg Val Glu Ile Trp His Gly Gly Ser
930 935 940
Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu Asp Asp Ala Gln Val
945 950 955 960
Val Cys Gln Gln Leu Gly Cys Gly Pro Ala Leu Lys Ala Phe Lys Glu
965 970 975
Ala Glu Phe Gly Gln Gly Thr Gly Pro Ile Trp Leu Asn Glu Val Lys
980 985 990
Cys Lys Gly Asn Glu Ser Ser Leu Trp Asp Cys Pro Ala Arg Arg Trp
995 1000 1005
Gly His Ser Glu Cys Gly His Lys Glu Asp Ala Ala Val Asn Cys
1010 1015 1020
Thr Asp Ile Ser Val Gln Lys Thr Pro Gln Lys Ala Thr Thr Gly
1025 1030 1035
Arg Ser Ser Arg Gln Ser Ser Phe Ile Ala Val Gly Ile Leu Gly
1040 1045 1050
Val Val Leu Leu Ala Ile Phe Val Ala Leu Phe Phe Leu Thr Lys
1055 1060 1065
Lys Arg Arg Gln Arg Gln Arg Leu Ala Val Ser Ser Arg Gly Glu
1070 1075 1080
Asn Leu Val His Gln Ile Gln Tyr Arg Glu Met Asn Ser Cys Leu
1085 1090 1095
Asn Ala Asp Asp Leu Asp Leu Met Asn Ser Ser Gly Leu Trp Val
1100 1105 1110
Leu Gly Gly Ser Ile Ala Gln Gly Phe Arg Ser Val Ala Ala Val
1115 1120 1125
Glu Ala Gln Thr Phe Tyr Phe Asp Lys Gln Leu Lys Lys Ser Lys
1130 1135 1140
Asn Val Ile Gly Ser Leu Asp Ala Tyr Asn Gly Gln Glu
1145 1150 1155
<210> SEQ ID NO 9
<211> LENGTH: 5193
<212> TYPE: DNA
<213> ORGANISM: sus scrofa
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/NM_214346
<309> DATABASE ENTRY DATE: 2004-08-05
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(5193)
<400> SEQUENCE: 9
atggacttcc tgctcctgct cctcctcctg gcttcatctg ctctagcagg cctggcctcg 60
tggacggttt ccagccccga gaccgtgcag ggcatcaagg gctcctgcct catcatcccc 120
tgcaccttcg gcttcccggc caacgtggag gtgccccatg gcatcacagc catctggtac 180
tatgactact caggcaagcg cctggtagtg agccactcca ggaacccaaa ggtggtggag 240
aaccacttcc aaggccgggc cctgctgttg gggcaggttg aacagaggac gtgcagcctg 300
ctgctgaagg acctgcagcc ccaggactcg ggctcctata acttccgctt tgagatcagc 360
gagggcaacc gctggtcaga tgtcaaaggc acagttgtca ccgtgacaga ggtgcccagc 420
gtgcccacca ttgccttgcc agccaagctg catgagggca tggaggtgga cttcaactgc 480
tccactccct atgtgtgccc gacggagccg gtcaacctac agtggcaagg ccaggatccc 540
acccgctccg tcacctccca cctccagaag cttgagccct cgggcaccag ccacatggag 600
accctgcaca tggccctgtc ctggcaggac catggccgga tcctgagctg ccaggtctca 660
gcagccgaac gcaggatgca gaaggagatt cacctccaag tgcagtatgc ccccaagggt 720
gtggagatcc ttttcagcca ctccggacgg aacgtccttc caggtgatct ggtcaccctc 780
agctgccagg tgaatagcag caaccctcag gtcagttccg tgcagtgggt caaggatggg 840
acgaagctca aagaccagaa acgtgtactg cagttgcgcc gggcagcctg ggctgatgct 900
ggcgtctaca cctgccaagc cgggaatgcc gtgggctctt cagtctcacc cccggtcagc 960
ctccacgtct tcatggctga ggtccaggta agccctgtgg gctccatcct ggagaaccag 1020
acggtgacgc tggcctgcaa tacacctaag gaagcgccca gcgagctgcg ctacagctgg 1080
tacaagaacc acgccctgct ggagggctct cacagccgca ccctccggct gcactcagtt 1140
accagggcgg attcgggctt ctacttctgc gaggtgcaga acgcccgggg cagagagcgc 1200
tctccccctg tcagcgtggt ggtcagccac ccacccctca ccccggacct aactgccttc 1260
ctggagacac aggcggggct ggtgggcatc ctccaatgct ctgtggtcag cgagccccca 1320
gctactctgg tgttgtcaca cgggggcctc atcttggcct ctacctccgg ggagggtgac 1380
cacagcccac gcttcagtgt cgcctctgcc cccaactccc tgcgcctgga gattcaagac 1440
ctggggccaa cagacagtgg ggaatacatg tgctcagcca gcagttctct tgggaatgcg 1500
tcctccaccc tggacttcca tgccaatgca gcccgcctcc tcatcagccc agcagcagag 1560
gtggtggaag ggcaggcggt gacactgagc tgcaggagca gcctgagcct gatgcctgac 1620
acccgttttt cctggtacct gaacggggcc ctgattctcg aggggcccag cagcagcctc 1680
ctgctcccag cagcctccag cacagatgcc ggctcatacc actgccgggc ccagaacagc 1740
cacagcacca gcgggccctc ctcacctgct gttctcaccg tgctctacgc cccacgccag 1800
cccgtgttca ctgcccagct ggaccctgat actgcaggag ctggggccgg acgccaaggc 1860
ctcctcttgt gccgtgtgga cagcgacccc ccagcccagc tgcagctgct ccacaggggc 1920
cgtgttgtgg cctcttctct gtcatggggg ggcggctgct gcacctgcgg aggctgtttc 1980
caccgcatga aggtcaccaa agcacccaac ctactgcgtg tagagatccg agacccggtg 2040
ctggaggatg agggtgtgta cctgtgcgag gccagcagcg ccctgggcaa cgcctccgcc 2100
tctgcaacct tggatgccca ggccactgtc ctggtcatca caccgtcaca cacgctgcag 2160
gaaggcattg aagccaacct gacttgcaac gtgagccgtg aagccagcgg ccctgccaac 2220
ttctcctggt tccgagatgg ggcgctatgg gcccagggcc ctctggacac cgtgacgctg 2280
ctacctgtgg ccagaactga tgctgccctc tatgcttgcc gcatcgtcac cgaggctggt 2340
gctggcctct ccacccctgt ggccctgaat gtgctctatc cccccgatcc tccaaagttg 2400
tcagccctcc tggacgtgga ccagggccac acggctgtgt tcgtctgtac tgtggacagt 2460
cgccctcttg cccagttggc cctgttccgt ggggaacacc tcctggccgc cagctcggca 2520
ctccggctcc cccctcgtgg ccgcctccag gccaaagcct cggccaactc cttgcagcta 2580
gaggtccgag acttgagcct tggggactct ggcagctacc actgtgaggc caccaacatc 2640
cttggatcag ccaacacttc tcttaccttc caggtccgag gagcctgggt ccgggtgtca 2700
ccgtcgcctg agctccagga gggccaggct gtggtcctga gctgccaggt acccataggg 2760
gtcctggagg ggacctcata tcgttggtat cgggatggcc agcccctcca ggagtccact 2820
tcggccacgc tccgttttgc agccataact ctgagccagg ctggagccta ccattgccaa 2880
gcccaagctc caggctcagc caccacggac ctggctgccc ctgtcagcct ccacgtgacc 2940
tacgcacctc gccaggccac actcaccacc ctgatggact caggcctcgg gcgactgggc 3000
ctccttctgt gccgtgtgaa cagtgaccct cctgcccagc tccgactgct ccatgggagc 3060
cgcctcgtgg cctctactct acaaggtgtg gaggagcttg caggcagctc tccccgccta 3120
caggtggcca cagcccccaa cacgctgcgc ctggagatcc acaacgcagt gctggaggat 3180
gaaggcgtct acacctgcga ggccaccaac accctgggtc agaccttggc ctccgccgcc 3240
ttcgatgccc aggctatgag agtgcaggtg tggcccaatg ccaccgtgca agaggggcag 3300
ctggtgaacc tgacctgcct tgtatggacc acgcacctgg cccagctcac ctacacgtgg 3360
taccgagacc agcagcagct cccaggtgct gcccactcca tcctcctgcc caatgtcact 3420
gtcacagatg ccgcctccta ccgctgtggc atattgatcc ctggccaggc actccgcctc 3480
tccagacctg tcgccctgga tgtcctctac gcaccccgca gactgcgcct gacccatctc 3540
ttggagagcc gtggtgggca gctggccgtg gtgctgtgca ctgtggacag tcgcccagct 3600
gcccagctga ccctcagcca tgctggccgc ctcctggcct cctcaaccgc agcctctgtc 3660
cccaacaccc tgcgcctgga gctgtgggag ccccggccca gtgatgaggg tctctacagc 3720
tgctcggccc gcagtcctct gggccaggcc aacacatccc tggagctgcg gctagagggc 3780
gtgcaggtgg cactggctcc atcggccact gtgccggagg gggcccctgt cacagtgacc 3840
tgtgaagacc ctgctgcccg cccacccact ctctatgtct ggtaccacaa cagccgttgg 3900
ctgcaggagg ggtcggctgc ctccctctcg tttccagcgg ctacacgggc tcacgcgggc 3960
gcctatacct gccaggtcca ggatgcccag ggcacacgca tctcccagcc cgcagcactg 4020
cacatcctct atgcccctcg ggatgctgtc ctttcctcct tctgggactc aagggccagc 4080
cctatggccg tggtacagtg cactgtggac agcgagccac ctgccgagat gaccctgtcc 4140
catgatggca aggtgctggc caccagccat ggggtccacg gcttagcagt ggggacaggc 4200
catgtccagg tggcccgcaa cgccctgcag ctgcgggtgc agaatgtgcc ctcacgtgac 4260
aaggacacct acgtctgcat ggaccgcaac tccttgggct cagtcagcac catggggcag 4320
ctgcagccag aaggtgtgca cgtggtagct gagccagggc tggatgtgcc tgaaggcaca 4380
gcgctgaacc tgagctgtcg cctccctagt ggccctgggc acataggcaa ctccaccttt 4440
gcttggttcc ggaacggtcg gcagctacac acagagtctg tgcccaccct taccttcacc 4500
catgtggccc gcgcccaagc tggcttgtac cactgccagg ctgagctccc cgccggggct 4560
gccacctctg ctccagtctt gctccgggtg ctctaccctc ccaagacgcc caccatgact 4620
gtttttgtgg agcccgaggg tggcatccag ggcattctgg actgccgagt ggacagtgag 4680
cccctagcca gcctgaccct ccacctgggc agtcggctgg tggcctccag ccagcctcag 4740
gctgcccctg ccaagccgca catccgcgtc tcagccagtc ccaatgcctt gcgagtggac 4800
atggaggagc tgaagcccag tgaccagggg gagtatgtgt gctcggcctc caatgccctg 4860
ggctctgcct ctgctgccac ctacttcgga accagagccc tgcatcgcct gcatctgttc 4920
cagcaccttc tctggttcct ggggctgctg gcgagcctcc tcttcctact gttgggcctg 4980
ggggtctggt acgcctggag acggggaaat ttttacaagc tgagaatggg cgaatattca 5040
gtagagatgg tatctcggaa ggaaaccacg cagatgtcca ctgaccagga agaagttact 5100
ggaatcggtg atgatgcggg ctctgtgaac caggcggcat ttgatcctgc ccacctctgt 5160
gaaaacacac agtctgtgaa aagcacagtc tga 5193
<210> SEQ ID NO 10
<211> LENGTH: 1730
<212> TYPE: PRT
<213> ORGANISM: sus scrofa
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/NP_999511
<309> DATABASE ENTRY DATE: 2004-08-05
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1730)
<400> SEQUENCE: 10
Met Asp Phe Leu Leu Leu Leu Leu Leu Leu Ala Ser Ser Ala Leu Ala
1 5 10 15
Gly Leu Ala Ser Trp Thr Val Ser Ser Pro Glu Thr Val Gln Gly Ile
20 25 30
Lys Gly Ser Cys Leu Ile Ile Pro Cys Thr Phe Gly Phe Pro Ala Asn
35 40 45
Val Glu Val Pro His Gly Ile Thr Ala Ile Trp Tyr Tyr Asp Tyr Ser
50 55 60
Gly Lys Arg Leu Val Val Ser His Ser Arg Asn Pro Lys Val Val Glu
65 70 75 80
Asn His Phe Gln Gly Arg Ala Leu Leu Leu Gly Gln Val Glu Gln Arg
85 90 95
Thr Cys Ser Leu Leu Leu Lys Asp Leu Gln Pro Gln Asp Ser Gly Ser
100 105 110
Tyr Asn Phe Arg Phe Glu Ile Ser Glu Gly Asn Arg Trp Ser Asp Val
115 120 125
Lys Gly Thr Val Val Thr Val Thr Glu Val Pro Ser Val Pro Thr Ile
130 135 140
Ala Leu Pro Ala Lys Leu His Glu Gly Met Glu Val Asp Phe Asn Cys
145 150 155 160
Ser Thr Pro Tyr Val Cys Pro Thr Glu Pro Val Asn Leu Gln Trp Gln
165 170 175
Gly Gln Asp Pro Thr Arg Ser Val Thr Ser His Leu Gln Lys Leu Glu
180 185 190
Pro Ser Gly Thr Ser His Met Glu Thr Leu His Met Ala Leu Ser Trp
195 200 205
Gln Asp His Gly Arg Ile Leu Ser Cys Gln Val Ser Ala Ala Glu Arg
210 215 220
Arg Met Gln Lys Glu Ile His Leu Gln Val Gln Tyr Ala Pro Lys Gly
225 230 235 240
Val Glu Ile Leu Phe Ser His Ser Gly Arg Asn Val Leu Pro Gly Asp
245 250 255
Leu Val Thr Leu Ser Cys Gln Val Asn Ser Ser Asn Pro Gln Val Ser
260 265 270
Ser Val Gln Trp Val Lys Asp Gly Thr Lys Leu Lys Asp Gln Lys Arg
275 280 285
Val Leu Gln Leu Arg Arg Ala Ala Trp Ala Asp Ala Gly Val Tyr Thr
290 295 300
Cys Gln Ala Gly Asn Ala Val Gly Ser Ser Val Ser Pro Pro Val Ser
305 310 315 320
Leu His Val Phe Met Ala Glu Val Gln Val Ser Pro Val Gly Ser Ile
325 330 335
Leu Glu Asn Gln Thr Val Thr Leu Ala Cys Asn Thr Pro Lys Glu Ala
340 345 350
Pro Ser Glu Leu Arg Tyr Ser Trp Tyr Lys Asn His Ala Leu Leu Glu
355 360 365
Gly Ser His Ser Arg Thr Leu Arg Leu His Ser Val Thr Arg Ala Asp
370 375 380
Ser Gly Phe Tyr Phe Cys Glu Val Gln Asn Ala Arg Gly Arg Glu Arg
385 390 395 400
Ser Pro Pro Val Ser Val Val Val Ser His Pro Pro Leu Thr Pro Asp
405 410 415
Leu Thr Ala Phe Leu Glu Thr Gln Ala Gly Leu Val Gly Ile Leu Gln
420 425 430
Cys Ser Val Val Ser Glu Pro Pro Ala Thr Leu Val Leu Ser His Gly
435 440 445
Gly Leu Ile Leu Ala Ser Thr Ser Gly Glu Gly Asp His Ser Pro Arg
450 455 460
Phe Ser Val Ala Ser Ala Pro Asn Ser Leu Arg Leu Glu Ile Gln Asp
465 470 475 480
Leu Gly Pro Thr Asp Ser Gly Glu Tyr Met Cys Ser Ala Ser Ser Ser
485 490 495
Leu Gly Asn Ala Ser Ser Thr Leu Asp Phe His Ala Asn Ala Ala Arg
500 505 510
Leu Leu Ile Ser Pro Ala Ala Glu Val Val Glu Gly Gln Ala Val Thr
515 520 525
Leu Ser Cys Arg Ser Ser Leu Ser Leu Met Pro Asp Thr Arg Phe Ser
530 535 540
Trp Tyr Leu Asn Gly Ala Leu Ile Leu Glu Gly Pro Ser Ser Ser Leu
545 550 555 560
Leu Leu Pro Ala Ala Ser Ser Thr Asp Ala Gly Ser Tyr His Cys Arg
565 570 575
Ala Gln Asn Ser His Ser Thr Ser Gly Pro Ser Ser Pro Ala Val Leu
580 585 590
Thr Val Leu Tyr Ala Pro Arg Gln Pro Val Phe Thr Ala Gln Leu Asp
595 600 605
Pro Asp Thr Ala Gly Ala Gly Ala Gly Arg Gln Gly Leu Leu Leu Cys
610 615 620
Arg Val Asp Ser Asp Pro Pro Ala Gln Leu Gln Leu Leu His Arg Gly
625 630 635 640
Arg Val Val Ala Ser Ser Leu Ser Trp Gly Gly Gly Cys Cys Thr Cys
645 650 655
Gly Gly Cys Phe His Arg Met Lys Val Thr Lys Ala Pro Asn Leu Leu
660 665 670
Arg Val Glu Ile Arg Asp Pro Val Leu Glu Asp Glu Gly Val Tyr Leu
675 680 685
Cys Glu Ala Ser Ser Ala Leu Gly Asn Ala Ser Ala Ser Ala Thr Leu
690 695 700
Asp Ala Gln Ala Thr Val Leu Val Ile Thr Pro Ser His Thr Leu Gln
705 710 715 720
Glu Gly Ile Glu Ala Asn Leu Thr Cys Asn Val Ser Arg Glu Ala Ser
725 730 735
Gly Pro Ala Asn Phe Ser Trp Phe Arg Asp Gly Ala Leu Trp Ala Gln
740 745 750
Gly Pro Leu Asp Thr Val Thr Leu Leu Pro Val Ala Arg Thr Asp Ala
755 760 765
Ala Leu Tyr Ala Cys Arg Ile Val Thr Glu Ala Gly Ala Gly Leu Ser
770 775 780
Thr Pro Val Ala Leu Asn Val Leu Tyr Pro Pro Asp Pro Pro Lys Leu
785 790 795 800
Ser Ala Leu Leu Asp Val Asp Gln Gly His Thr Ala Val Phe Val Cys
805 810 815
Thr Val Asp Ser Arg Pro Leu Ala Gln Leu Ala Leu Phe Arg Gly Glu
820 825 830
His Leu Leu Ala Ala Ser Ser Ala Leu Arg Leu Pro Pro Arg Gly Arg
835 840 845
Leu Gln Ala Lys Ala Ser Ala Asn Ser Leu Gln Leu Glu Val Arg Asp
850 855 860
Leu Ser Leu Gly Asp Ser Gly Ser Tyr His Cys Glu Ala Thr Asn Ile
865 870 875 880
Leu Gly Ser Ala Asn Thr Ser Leu Thr Phe Gln Val Arg Gly Ala Trp
885 890 895
Val Arg Val Ser Pro Ser Pro Glu Leu Gln Glu Gly Gln Ala Val Val
900 905 910
Leu Ser Cys Gln Val Pro Ile Gly Val Leu Glu Gly Thr Ser Tyr Arg
915 920 925
Trp Tyr Arg Asp Gly Gln Pro Leu Gln Glu Ser Thr Ser Ala Thr Leu
930 935 940
Arg Phe Ala Ala Ile Thr Leu Ser Gln Ala Gly Ala Tyr His Cys Gln
945 950 955 960
Ala Gln Ala Pro Gly Ser Ala Thr Thr Asp Leu Ala Ala Pro Val Ser
965 970 975
Leu His Val Thr Tyr Ala Pro Arg Gln Ala Thr Leu Thr Thr Leu Met
980 985 990
Asp Ser Gly Leu Gly Arg Leu Gly Leu Leu Leu Cys Arg Val Asn Ser
995 1000 1005
Asp Pro Pro Ala Gln Leu Arg Leu Leu His Gly Ser Arg Leu Val
1010 1015 1020
Ala Ser Thr Leu Gln Gly Val Glu Glu Leu Ala Gly Ser Ser Pro
1025 1030 1035
Arg Leu Gln Val Ala Thr Ala Pro Asn Thr Leu Arg Leu Glu Ile
1040 1045 1050
His Asn Ala Val Leu Glu Asp Glu Gly Val Tyr Thr Cys Glu Ala
1055 1060 1065
Thr Asn Thr Leu Gly Gln Thr Leu Ala Ser Ala Ala Phe Asp Ala
1070 1075 1080
Gln Ala Met Arg Val Gln Val Trp Pro Asn Ala Thr Val Gln Glu
1085 1090 1095
Gly Gln Leu Val Asn Leu Thr Cys Leu Val Trp Thr Thr His Leu
1100 1105 1110
Ala Gln Leu Thr Tyr Thr Trp Tyr Arg Asp Gln Gln Gln Leu Pro
1115 1120 1125
Gly Ala Ala His Ser Ile Leu Leu Pro Asn Val Thr Val Thr Asp
1130 1135 1140
Ala Ala Ser Tyr Arg Cys Gly Ile Leu Ile Pro Gly Gln Ala Leu
1145 1150 1155
Arg Leu Ser Arg Pro Val Ala Leu Asp Val Leu Tyr Ala Pro Arg
1160 1165 1170
Arg Leu Arg Leu Thr His Leu Leu Glu Ser Arg Gly Gly Gln Leu
1175 1180 1185
Ala Val Val Leu Cys Thr Val Asp Ser Arg Pro Ala Ala Gln Leu
1190 1195 1200
Thr Leu Ser His Ala Gly Arg Leu Leu Ala Ser Ser Thr Ala Ala
1205 1210 1215
Ser Val Pro Asn Thr Leu Arg Leu Glu Leu Trp Glu Pro Arg Pro
1220 1225 1230
Ser Asp Glu Gly Leu Tyr Ser Cys Ser Ala Arg Ser Pro Leu Gly
1235 1240 1245
Gln Ala Asn Thr Ser Leu Glu Leu Arg Leu Glu Gly Val Gln Val
1250 1255 1260
Ala Leu Ala Pro Ser Ala Thr Val Pro Glu Gly Ala Pro Val Thr
1265 1270 1275
Val Thr Cys Glu Asp Pro Ala Ala Arg Pro Pro Thr Leu Tyr Val
1280 1285 1290
Trp Tyr His Asn Ser Arg Trp Leu Gln Glu Gly Ser Ala Ala Ser
1295 1300 1305
Leu Ser Phe Pro Ala Ala Thr Arg Ala His Ala Gly Ala Tyr Thr
1310 1315 1320
Cys Gln Val Gln Asp Ala Gln Gly Thr Arg Ile Ser Gln Pro Ala
1325 1330 1335
Ala Leu His Ile Leu Tyr Ala Pro Arg Asp Ala Val Leu Ser Ser
1340 1345 1350
Phe Trp Asp Ser Arg Ala Ser Pro Met Ala Val Val Gln Cys Thr
1355 1360 1365
Val Asp Ser Glu Pro Pro Ala Glu Met Thr Leu Ser His Asp Gly
1370 1375 1380
Lys Val Leu Ala Thr Ser His Gly Val His Gly Leu Ala Val Gly
1385 1390 1395
Thr Gly His Val Gln Val Ala Arg Asn Ala Leu Gln Leu Arg Val
1400 1405 1410
Gln Asn Val Pro Ser Arg Asp Lys Asp Thr Tyr Val Cys Met Asp
1415 1420 1425
Arg Asn Ser Leu Gly Ser Val Ser Thr Met Gly Gln Leu Gln Pro
1430 1435 1440
Glu Gly Val His Val Val Ala Glu Pro Gly Leu Asp Val Pro Glu
1445 1450 1455
Gly Thr Ala Leu Asn Leu Ser Cys Arg Leu Pro Ser Gly Pro Gly
1460 1465 1470
His Ile Gly Asn Ser Thr Phe Ala Trp Phe Arg Asn Gly Arg Gln
1475 1480 1485
Leu His Thr Glu Ser Val Pro Thr Leu Thr Phe Thr His Val Ala
1490 1495 1500
Arg Ala Gln Ala Gly Leu Tyr His Cys Gln Ala Glu Leu Pro Ala
1505 1510 1515
Gly Ala Ala Thr Ser Ala Pro Val Leu Leu Arg Val Leu Tyr Pro
1520 1525 1530
Pro Lys Thr Pro Thr Met Thr Val Phe Val Glu Pro Glu Gly Gly
1535 1540 1545
Ile Gln Gly Ile Leu Asp Cys Arg Val Asp Ser Glu Pro Leu Ala
1550 1555 1560
Ser Leu Thr Leu His Leu Gly Ser Arg Leu Val Ala Ser Ser Gln
1565 1570 1575
Pro Gln Ala Ala Pro Ala Lys Pro His Ile Arg Val Ser Ala Ser
1580 1585 1590
Pro Asn Ala Leu Arg Val Asp Met Glu Glu Leu Lys Pro Ser Asp
1595 1600 1605
Gln Gly Glu Tyr Val Cys Ser Ala Ser Asn Ala Leu Gly Ser Ala
1610 1615 1620
Ser Ala Ala Thr Tyr Phe Gly Thr Arg Ala Leu His Arg Leu His
1625 1630 1635
Leu Phe Gln His Leu Leu Trp Phe Leu Gly Leu Leu Ala Ser Leu
1640 1645 1650
Leu Phe Leu Leu Leu Gly Leu Gly Val Trp Tyr Ala Trp Arg Arg
1655 1660 1665
Gly Asn Phe Tyr Lys Leu Arg Met Gly Glu Tyr Ser Val Glu Met
1670 1675 1680
Val Ser Arg Lys Glu Thr Thr Gln Met Ser Thr Asp Gln Glu Glu
1685 1690 1695
Val Thr Gly Ile Gly Asp Asp Ala Gly Ser Val Asn Gln Ala Ala
1700 1705 1710
Phe Asp Pro Ala His Leu Cys Glu Asn Thr Gln Ser Val Lys Ser
1715 1720 1725
Thr Val
1730
<210> SEQ ID NO 11
<211> LENGTH: 6387
<212> TYPE: DNA
<213> ORGANISM: mus musculus
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/NM_011426
<309> DATABASE ENTRY DATE: 2000-01-25
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(6387)
<400> SEQUENCE: 11
agacaagatt aggcctagag taagtctatg aaacacagag aaaggggaca gcataggggt 60
taagaaatga ggtctttcaa aatctcaggg ggcaatgagg agttttttga gagaggaagg 120
actctttaaa ggaagttgaa ggaggattct gtgaacttga gaccaccctg agctgccaag 180
ttgagaactt tgtctacaaa caagccaggc agcctcagcg tgtgctcagt ccgacttgta 240
gctggagagg caggagacca atttccggtg cttacggtgc ttgctggatg ccctggagta 300
agtgacaggg tctcactgga ctccaggttc tgttggtttg agtaatagga ggcggcaggg 360
gagaagtgaa gagagacatg cactgctgat ctgccttgag gctgtgtcct taaggggtgg 420
agccaagggg cacagaagac tctctgggac atgccaccaa gtgagagcat ttccaatcac 480
tccctgagcc aggaacaggg gcttctggtt ccctgctggt ggctgccaca gcagtccttc 540
ctgttgggtt gaccaacaca gcaggtgaga taaaccctat agacttgggc cctggagtgc 600
tccaggcagt ctctgtgtgc ctacccaccc ggcttcccta ggcacctgaa tgcacctggg 660
cactgggatg tgtgtcctgt tctccctgct cctgctggcc tctgtcttct cactaggcca 720
gaccacatgg ggtgtctcca gtcccaagaa tgtgcagggc ttgtcgggat cctgcctgct 780
cattccctgc atcttcagct accctgccga tgtcccagtg tccaatggca tcacagccat 840
ctggtactat gactactcgg gcaagcggca ggtggtaatc cactcagggg accccaagct 900
ggtggacaag cgtttcaggg gtcgagctga actgatgggg aacatggacc acaaggtgtg 960
caacctgttg ctcaaagact tgaagcctga agactctggc acctacaact tccgctttga 1020
gatcagtgat agcaaccgct ggttagatgt caaaggcacc acggtcactg tgacaacgga 1080
tcccagcccc cccactatta ccattcctga ggagctgcgt gaaggcatgg agaggaactt 1140
caactgttcc acaccctacc tgtgcctgca ggagaagcaa gtcagcctgc agtggcgagg 1200
ccaggacccc acccactctg tcacctccag cttccagagc ctcgagccca ctggcgtcta 1260
tcaccagacg accctacata tggccctatc ctggcaggac cacggtcgga ccctgctctg 1320
ccagttctca ttgggcgcac acagtagtcg gaaagaggtt tacctgcaag tgccacatgc 1380
ccccaaaggt gtggagatcc tcctcagctc ctcagggagg aacatccttc ccggggatcc 1440
agtcacactc acctgcagag tgaacagcag ctatcctgct gtcagtgccg tgcagtgggc 1500
cagggacgga gtgaacctcg gagtcacggg acatgtgctt cggctgttct cagcagcctg 1560
gaatgattct ggggcctaca cctgccaagc aacaaatgat atgggctctc tggtgtcatc 1620
cccgctcagc ctccatgttt ttatggctga agtcaaaatg aaccccgcag ggcccgtctt 1680
ggaaaatgag acagtgactc tgctctgtag cacgccgaag gaggctcccc aggagctccg 1740
ctatagctgg tacaagaacc acattctcct ggaagatgcc catgcctcaa ccttgcacct 1800
gcctgcagtc accagggctg atactggctt ctacttctgt gaagtgcaga atgcccaggg 1860
cagtgagcgc tccagtccat tgagtgtggt ggtcagatat ccacccctta ctccagacct 1920
gaccaccttc ctggagacac aggccggact tgtgggcatc ttgcattgct ccgtggtcag 1980
tgagcccctg gctactgtgg tgctgtcaca cggaggcctc acgttggcct ccaactctgg 2040
agaaaatgac ttcaaccccc gattcaggat ctcctctgcc cccaactccc tgcgcctaga 2100
aatccgagac ttgcagccag cagacagcgg agagtacaca tgcttagctg tcaactccct 2160
tggaaactca acgtccagcc tagacttcta tgctaatgtg gcccgactcc tcatcaaccc 2220
ttcagcagag gttgtggaag ggcaggcggt gaccctgagc tgcaggagtg gcctgagccc 2280
agctcctgac actcgcttct cctggtacct gaacggagct ctacttctgg aaggatccag 2340
cagcagcctc ctgcttcctg cggcttccag cactgatgcg ggctcatact actgtaggac 2400
gcaggctggc cccaacacca gcggcccctc cctgcctact gtcctcactg tgttctatcc 2460
cccaagaaag cccacattca ctgccaggct ggatttggat acctctggag tcggggatgg 2520
acgacggggc atcctcttgt gccacgtaga cagcgatccc ccagcccagc tacggcttct 2580
ccacaaaggc catgttgtgg ccacttctct gccatcaagg tgtgggagct gttcccagcg 2640
cacaaaagtc agcagaacct ccaactcact gcacgtggag atccagaagc ctgtattaga 2700
ggatgagggc gtgtacctgt gtgaggctag caacacattg ggcaactcct cagccgcagc 2760
ctctttcaat gctaaggcca ctgtactggt catcacaccg tcaaatacac tgcgtgaagg 2820
cacagaggcc aacctaactt gcaacgtgaa ccaggaggtt gctgtcagcc ctgccaactt 2880
ctcctggttc cggaatggag tgctgtggac ccagggatca ctggagactg tgaggctgca 2940
gcctgtggcc agaactgatg ctgctgtcta tgcctgccgc ctcctcaccg aggatggggc 3000
tcagctctcg gctcctgtgg tcctaagtgt gctgtatgcc ccagaccctc caaagctgtc 3060
agccctccta gatgtgggtc agggccacat ggccgtgttc atctgcactg tggacagcta 3120
tcccctggct cacctgtctc tgttccgtgg ggaccatctc ctggccacca acttggaacc 3180
ccagcgtccc tcccatggca ggatccaggc caaggccaca gccaactccc tgcagctaga 3240
ggtccgagaa ctaggtcttg tggactctgg aaactaccac tgtgaagcca ccaatattct 3300
tgggtcagcc aacagttcac tcttcttcca ggtcagagga gcctgggtcc aggtttcacc 3360
atcacctgag ctccgggagg gccaggctgt ggtcctgagc tgccaggtgc ccacaggagt 3420
ctctgagggg acctcataca gctggtatca ggatggccgc cccctccagg agtcaacctc 3480
atctacactc cgcattgcag ccataagtct gaggcaagct ggtgcctacc attgccaagc 3540
tcaggcccca gacacagcta ttgccagcct ggctgcccct gtcagcctcc atgtgtccta 3600
taccccacgt catgttacac tcagtgccct gctgagcacg gaccctgagc gactaggcca 3660
cctggtgtgc agtgtacaaa gtgaccctcc agcgcagctg caactgtttc accggaatcg 3720
cctcgtggcc tctaccctac aaggcgcgga cgaattggca ggcagtaatc cccggctgca 3780
tgtgactgtg ctccccaatg agctgcgcct gcagatccac tttccagagc tggaggatga 3840
cgggacctat acatgcgaag ccagcaacac actgggccag gcctcggctg cagctgactt 3900
cgatgcccag gctgtgcgag tgactgtgtg gcccaatgcc actgtgcaag aggggcagca 3960
ggtgaacctg acctgcttgg tgtggagcac ccaccaggac tcactcagct acacatggta 4020
caagggcggg caacaactcc ttggtgccag atccatcacc ctgcccagtg ttaaggtttt 4080
ggatgctacc tcctaccgct gtggtgtggg gctccccggc cacgcacccc atctctccag 4140
acccgtgacc ctggatgtcc tccatgctcc ccgaaacctg cggctgacct acctcctaga 4200
gacccagggc aggcagctgg ccctggtact gtgtacggtg gatagtcgtc cacctgccca 4260
gctaactctc agccatggtg accagcttgt agcctcctca actgaagcct ctgtccccaa 4320
caccctgcgc ctagagcttc aggatccaag gcctagtaat gaggggctct atagctgctc 4380
tgcccacagc ccattgggca aggccaacac gtccctggaa cttctgctgg aaggtgtccg 4440
agtgaaaatg aatccctctg gtagtgtacc cgagggagag cctgtcacag tgacctgcga 4500
ggaccctgct gccctctcat ccgccctcta tgcctggttt cacaatggcc attggcttca 4560
ggagggaccg gcttcctcac tccagttcct ggtgactaca cgggctcacg ctggtgctta 4620
cttttgccag gtgcatgata cacaaggcac acggagctcc agacctgcca gcctgcaaat 4680
tctctatgcc ccccgggatg ctgtcctgtc ttcctttcga gactcaagga ccaggctcat 4740
ggtcgtgatt cagtgcaccg tggacagtga gccacctgct gagatggtcc tatcccacaa 4800
tggcaaggtg ctagctgcca gccacgagcg tcacagctca gcatcaggga taggccacat 4860
ccaggtagcc cgaaatgctc ttcgactaca agtgcaagat gtgactctgg gtgatggcaa 4920
cacctatgtt tgcacagccc agaatacact gggctccatc agtaccaccc agaggcttct 4980
gacggagact gatatacgtg tgacagctga gccaggcttg gatgtgccag agggcacagc 5040
tctgaactta agctgcctcc tccctggtgg ctctgggccc acgggcaact cttccttcac 5100
gtggttctgg aatcgccacc gactacattc agctcctgtg cccacactct ccttcacccc 5160
tgtggtccgg gctcaggctg ggctgtacca ctgcagggct gatctcccca ccggggccac 5220
tacctctgct ccagttatgc tccgtgtcct ctatcccccc aagacgccca ctctcatagt 5280
gtttgtggag cctcagggtg gccaccaggg catcctcgac tgtcgagtgg acagtgagcc 5340
cctggccatc ctcactcttc accggggcag tcaactagta gcctccaacc aacttcacga 5400
tgctcccacc aagccccaca tccgagtcac tgctcctccc aatgccttga gagtggacat 5460
agaggagctc ggccctagca atcaagggga gtatgtgtgc actgcctcca acactctggg 5520
ctctgcctca gcctctgcct actttgggac cagagctctg caccaactgc agctgttcca 5580
gaggctgctc tgggtcctgg gatttctggc aggcttcctg tgcctgctgc tgggtctggt 5640
ggcctatcac acctggagaa agaagagttc taccaagctg aatgaggatg agaattcagc 5700
agagatggcc actaagaaaa atactatcca ggaggaagtg gttgctgctc tctgacaact 5760
caggtgctgt gaacaagatc ctgcctacct ctgtataagc agtacagaga catctggctt 5820
tcctgacctg cccgacttgc cttccaagcc tcttgatcct aagaaaaatg gacgaaggga 5880
ggtttggggt tggaggtcaa cctgccgcct ccagggctct gagacggact cagccatgtt 5940
gcccacgtct ctctgtgtgg ttttcctctg tatccctttg cctttctctt caaagctcac 6000
cttggacttt cttggtgggt tagagcaaca tccagtttct cacagacttt ctaagacggt 6060
ctgtaccagc caggatatca gtcaggttgc tctaacagag actcaataca gtgaccacag 6120
catgacaggg tcttagtttt ccctcctggc ctggttatgt tgttgtggta tcagaatcct 6180
tcttgcttga ttttctccat tccccaagtg ttgcctttga ttatgaagct caggtaactg 6240
cagtgcccat ggaccctaca gggagaagga agagtgaagg gaagacatac ccatccccat 6300
ggtccatgga ctgtgtgtgc aattgcaccc cacccaactt ctcatccgct agaaactggt 6360
cacataaaca taccatgctg aaaggga 6387
<210> SEQ ID NO 12
<211> LENGTH: 1695
<212> TYPE: PRT
<213> ORGANISM: mus musculus
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept
<309> DATABASE ENTRY DATE: 2000-01-25
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1695)
<400> SEQUENCE: 12
Met Cys Val Leu Phe Ser Leu Leu Leu Leu Ala Ser Val Phe Ser Leu
1 5 10 15
Gly Gln Thr Thr Trp Gly Val Ser Ser Pro Lys Asn Val Gln Gly Leu
20 25 30
Ser Gly Ser Cys Leu Leu Ile Pro Cys Ile Phe Ser Tyr Pro Ala Asp
35 40 45
Val Pro Val Ser Asn Gly Ile Thr Ala Ile Trp Tyr Tyr Asp Tyr Ser
50 55 60
Gly Lys Arg Gln Val Val Ile His Ser Gly Asp Pro Lys Leu Val Asp
65 70 75 80
Lys Arg Phe Arg Gly Arg Ala Glu Leu Met Gly Asn Met Asp His Lys
85 90 95
Val Cys Asn Leu Leu Leu Lys Asp Leu Lys Pro Glu Asp Ser Gly Thr
100 105 110
Tyr Asn Phe Arg Phe Glu Ile Ser Asp Ser Asn Arg Trp Leu Asp Val
115 120 125
Lys Gly Thr Thr Val Thr Val Thr Thr Asp Pro Ser Pro Pro Thr Ile
130 135 140
Thr Ile Pro Glu Glu Leu Arg Glu Gly Met Glu Arg Asn Phe Asn Cys
145 150 155 160
Ser Thr Pro Tyr Leu Cys Leu Gln Glu Lys Gln Val Ser Leu Gln Trp
165 170 175
Arg Gly Gln Asp Pro Thr His Ser Val Thr Ser Ser Phe Gln Ser Leu
180 185 190
Glu Pro Thr Gly Val Tyr His Gln Thr Thr Leu His Met Ala Leu Ser
195 200 205
Trp Gln Asp His Gly Arg Thr Leu Leu Cys Gln Phe Ser Leu Gly Ala
210 215 220
His Ser Ser Arg Lys Glu Val Tyr Leu Gln Val Pro His Ala Pro Lys
225 230 235 240
Gly Val Glu Ile Leu Leu Ser Ser Ser Gly Arg Asn Ile Leu Pro Gly
245 250 255
Asp Pro Val Thr Leu Thr Cys Arg Val Asn Ser Ser Tyr Pro Ala Val
260 265 270
Ser Ala Val Gln Trp Ala Arg Asp Gly Val Asn Leu Gly Val Thr Gly
275 280 285
His Val Leu Arg Leu Phe Ser Ala Ala Trp Asn Asp Ser Gly Ala Tyr
290 295 300
Thr Cys Gln Ala Thr Asn Asp Met Gly Ser Leu Val Ser Ser Pro Leu
305 310 315 320
Ser Leu His Val Phe Met Ala Glu Val Lys Met Asn Pro Ala Gly Pro
325 330 335
Val Leu Glu Asn Glu Thr Val Thr Leu Leu Cys Ser Thr Pro Lys Glu
340 345 350
Ala Pro Gln Glu Leu Arg Tyr Ser Trp Tyr Lys Asn His Ile Leu Leu
355 360 365
Glu Asp Ala His Ala Ser Thr Leu His Leu Pro Ala Val Thr Arg Ala
370 375 380
Asp Thr Gly Phe Tyr Phe Cys Glu Val Gln Asn Ala Gln Gly Ser Glu
385 390 395 400
Arg Ser Ser Pro Leu Ser Val Val Val Arg Tyr Pro Pro Leu Thr Pro
405 410 415
Asp Leu Thr Thr Phe Leu Glu Thr Gln Ala Gly Leu Val Gly Ile Leu
420 425 430
His Cys Ser Val Val Ser Glu Pro Leu Ala Thr Val Val Leu Ser His
435 440 445
Gly Gly Leu Thr Leu Ala Ser Asn Ser Gly Glu Asn Asp Phe Asn Pro
450 455 460
Arg Phe Arg Ile Ser Ser Ala Pro Asn Ser Leu Arg Leu Glu Ile Arg
465 470 475 480
Asp Leu Gln Pro Ala Asp Ser Gly Glu Tyr Thr Cys Leu Ala Val Asn
485 490 495
Ser Leu Gly Asn Ser Thr Ser Ser Leu Asp Phe Tyr Ala Asn Val Ala
500 505 510
Arg Leu Leu Ile Asn Pro Ser Ala Glu Val Val Glu Gly Gln Ala Val
515 520 525
Thr Leu Ser Cys Arg Ser Gly Leu Ser Pro Ala Pro Asp Thr Arg Phe
530 535 540
Ser Trp Tyr Leu Asn Gly Ala Leu Leu Leu Glu Gly Ser Ser Ser Ser
545 550 555 560
Leu Leu Leu Pro Ala Ala Ser Ser Thr Asp Ala Gly Ser Tyr Tyr Cys
565 570 575
Arg Thr Gln Ala Gly Pro Asn Thr Ser Gly Pro Ser Leu Pro Thr Val
580 585 590
Leu Thr Val Phe Tyr Pro Pro Arg Lys Pro Thr Phe Thr Ala Arg Leu
595 600 605
Asp Leu Asp Thr Ser Gly Val Gly Asp Gly Arg Arg Gly Ile Leu Leu
610 615 620
Cys His Val Asp Ser Asp Pro Pro Ala Gln Leu Arg Leu Leu His Lys
625 630 635 640
Gly His Val Val Ala Thr Ser Leu Pro Ser Arg Cys Gly Ser Cys Ser
645 650 655
Gln Arg Thr Lys Val Ser Arg Thr Ser Asn Ser Leu His Val Glu Ile
660 665 670
Gln Lys Pro Val Leu Glu Asp Glu Gly Val Tyr Leu Cys Glu Ala Ser
675 680 685
Asn Thr Leu Gly Asn Ser Ser Ala Ala Ala Ser Phe Asn Ala Lys Ala
690 695 700
Thr Val Leu Val Ile Thr Pro Ser Asn Thr Leu Arg Glu Gly Thr Glu
705 710 715 720
Ala Asn Leu Thr Cys Asn Val Asn Gln Glu Val Ala Val Ser Pro Ala
725 730 735
Asn Phe Ser Trp Phe Arg Asn Gly Val Leu Trp Thr Gln Gly Ser Leu
740 745 750
Glu Thr Val Arg Leu Gln Pro Val Ala Arg Thr Asp Ala Ala Val Tyr
755 760 765
Ala Cys Arg Leu Leu Thr Glu Asp Gly Ala Gln Leu Ser Ala Pro Val
770 775 780
Val Leu Ser Val Leu Tyr Ala Pro Asp Pro Pro Lys Leu Ser Ala Leu
785 790 795 800
Leu Asp Val Gly Gln Gly His Met Ala Val Phe Ile Cys Thr Val Asp
805 810 815
Ser Tyr Pro Leu Ala His Leu Ser Leu Phe Arg Gly Asp His Leu Leu
820 825 830
Ala Thr Asn Leu Glu Pro Gln Arg Pro Ser His Gly Arg Ile Gln Ala
835 840 845
Lys Ala Thr Ala Asn Ser Leu Gln Leu Glu Val Arg Glu Leu Gly Leu
850 855 860
Val Asp Ser Gly Asn Tyr His Cys Glu Ala Thr Asn Ile Leu Gly Ser
865 870 875 880
Ala Asn Ser Ser Leu Phe Phe Gln Val Arg Gly Ala Trp Val Gln Val
885 890 895
Ser Pro Ser Pro Glu Leu Arg Glu Gly Gln Ala Val Val Leu Ser Cys
900 905 910
Gln Val Pro Thr Gly Val Ser Glu Gly Thr Ser Tyr Ser Trp Tyr Gln
915 920 925
Asp Gly Arg Pro Leu Gln Glu Ser Thr Ser Ser Thr Leu Arg Ile Ala
930 935 940
Ala Ile Ser Leu Arg Gln Ala Gly Ala Tyr His Cys Gln Ala Gln Ala
945 950 955 960
Pro Asp Thr Ala Ile Ala Ser Leu Ala Ala Pro Val Ser Leu His Val
965 970 975
Ser Tyr Thr Pro Arg His Val Thr Leu Ser Ala Leu Leu Ser Thr Asp
980 985 990
Pro Glu Arg Leu Gly His Leu Val Cys Ser Val Gln Ser Asp Pro Pro
995 1000 1005
Ala Gln Leu Gln Leu Phe His Arg Asn Arg Leu Val Ala Ser Thr
1010 1015 1020
Leu Gln Gly Ala Asp Glu Leu Ala Gly Ser Asn Pro Arg Leu His
1025 1030 1035
Val Thr Val Leu Pro Asn Glu Leu Arg Leu Gln Ile His Phe Pro
1040 1045 1050
Glu Leu Glu Asp Asp Gly Thr Tyr Thr Cys Glu Ala Ser Asn Thr
1055 1060 1065
Leu Gly Gln Ala Ser Ala Ala Ala Asp Phe Asp Ala Gln Ala Val
1070 1075 1080
Arg Val Thr Val Trp Pro Asn Ala Thr Val Gln Glu Gly Gln Gln
1085 1090 1095
Val Asn Leu Thr Cys Leu Val Trp Ser Thr His Gln Asp Ser Leu
1100 1105 1110
Ser Tyr Thr Trp Tyr Lys Gly Gly Gln Gln Leu Leu Gly Ala Arg
1115 1120 1125
Ser Ile Thr Leu Pro Ser Val Lys Val Leu Asp Ala Thr Ser Tyr
1130 1135 1140
Arg Cys Gly Val Gly Leu Pro Gly His Ala Pro His Leu Ser Arg
1145 1150 1155
Pro Val Thr Leu Asp Val Leu His Ala Pro Arg Asn Leu Arg Leu
1160 1165 1170
Thr Tyr Leu Leu Glu Thr Gln Gly Arg Gln Leu Ala Leu Val Leu
1175 1180 1185
Cys Thr Val Asp Ser Arg Pro Pro Ala Gln Leu Thr Leu Ser His
1190 1195 1200
Gly Asp Gln Leu Val Ala Ser Ser Thr Glu Ala Ser Val Pro Asn
1205 1210 1215
Thr Leu Arg Leu Glu Leu Gln Asp Pro Arg Pro Ser Asn Glu Gly
1220 1225 1230
Leu Tyr Ser Cys Ser Ala His Ser Pro Leu Gly Lys Ala Asn Thr
1235 1240 1245
Ser Leu Glu Leu Leu Leu Glu Gly Val Arg Val Lys Met Asn Pro
1250 1255 1260
Ser Gly Ser Val Pro Glu Gly Glu Pro Val Thr Val Thr Cys Glu
1265 1270 1275
Asp Pro Ala Ala Leu Ser Ser Ala Leu Tyr Ala Trp Phe His Asn
1280 1285 1290
Gly His Trp Leu Gln Glu Gly Pro Ala Ser Ser Leu Gln Phe Leu
1295 1300 1305
Val Thr Thr Arg Ala His Ala Gly Ala Tyr Phe Cys Gln Val His
1310 1315 1320
Asp Thr Gln Gly Thr Arg Ser Ser Arg Pro Ala Ser Leu Gln Ile
1325 1330 1335
Leu Tyr Ala Pro Arg Asp Ala Val Leu Ser Ser Phe Arg Asp Ser
1340 1345 1350
Arg Thr Arg Leu Met Val Val Ile Gln Cys Thr Val Asp Ser Glu
1355 1360 1365
Pro Pro Ala Glu Met Val Leu Ser His Asn Gly Lys Val Leu Ala
1370 1375 1380
Ala Ser His Glu Arg His Ser Ser Ala Ser Gly Ile Gly His Ile
1385 1390 1395
Gln Val Ala Arg Asn Ala Leu Arg Leu Gln Val Gln Asp Val Thr
1400 1405 1410
Leu Gly Asp Gly Asn Thr Tyr Val Cys Thr Ala Gln Asn Thr Leu
1415 1420 1425
Gly Ser Ile Ser Thr Thr Gln Arg Leu Leu Thr Glu Thr Asp Ile
1430 1435 1440
Arg Val Thr Ala Glu Pro Gly Leu Asp Val Pro Glu Gly Thr Ala
1445 1450 1455
Leu Asn Leu Ser Cys Leu Leu Pro Gly Gly Ser Gly Pro Thr Gly
1460 1465 1470
Asn Ser Ser Phe Thr Trp Phe Trp Asn Arg His Arg Leu His Ser
1475 1480 1485
Ala Pro Val Pro Thr Leu Ser Phe Thr Pro Val Val Arg Ala Gln
1490 1495 1500
Ala Gly Leu Tyr His Cys Arg Ala Asp Leu Pro Thr Gly Ala Thr
1505 1510 1515
Thr Ser Ala Pro Val Met Leu Arg Val Leu Tyr Pro Pro Lys Thr
1520 1525 1530
Pro Thr Leu Ile Val Phe Val Glu Pro Gln Gly Gly His Gln Gly
1535 1540 1545
Ile Leu Asp Cys Arg Val Asp Ser Glu Pro Leu Ala Ile Leu Thr
1550 1555 1560
Leu His Arg Gly Ser Gln Leu Val Ala Ser Asn Gln Leu His Asp
1565 1570 1575
Ala Pro Thr Lys Pro His Ile Arg Val Thr Ala Pro Pro Asn Ala
1580 1585 1590
Leu Arg Val Asp Ile Glu Glu Leu Gly Pro Ser Asn Gln Gly Glu
1595 1600 1605
Tyr Val Cys Thr Ala Ser Asn Thr Leu Gly Ser Ala Ser Ala Ser
1610 1615 1620
Ala Tyr Phe Gly Thr Arg Ala Leu His Gln Leu Gln Leu Phe Gln
1625 1630 1635
Arg Leu Leu Trp Val Leu Gly Phe Leu Ala Gly Phe Leu Cys Leu
1640 1645 1650
Leu Leu Gly Leu Val Ala Tyr His Thr Trp Arg Lys Lys Ser Ser
1655 1660 1665
Thr Lys Leu Asn Glu Asp Glu Asn Ser Ala Glu Met Ala Thr Lys
1670 1675 1680
Lys Asn Thr Ile Gln Glu Glu Val Val Ala Ala Leu
1685 1690 1695
<210> SEQ ID NO 13
<211> LENGTH: 6736
<212> TYPE: DNA
<213> ORGANISM: homo sapiens
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenBank/NM_023068
<309> DATABASE ENTRY DATE: 2001-02-13
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(6736)
<400> SEQUENCE: 13
atgggcttct tgcccaagct tctcctcctg gcctcattct tcccagcagg ccaggcctca 60
tggggcgtct ccagtcccca ggacgtgcag ggtgtgaagg ggtcttgcct gcttatcccc 120
tgcatcttca gcttccctgc cgacgtggag gtgcccgacg gcatcacggc catctggtac 180
tacgactact cgggccagcg gcaggtggtg agccactcgg cggaccccaa gctggtggag 240
gcccgcttcc gcggccgcac cgagttcatg gggaaccccg agcacagggt gtgcaacctg 300
ctgctgaagg acctgcagcc cgaggactct ggttcctaca acttccgctt cgagatcagt 360
gaggtcaacc gctggtcaga tgtgaaaggc accttggtca cagtaacaga ggagcccagg 420
gtgcccacca ttgcctcccc ggtggagctt ctcgagggca cagaggtgga cttcaactgc 480
tccactccct acgtatgcct gcaggagcag gtcagactgc agtggcaagg ccaggaccct 540
gctcgctctg tcaccttcaa cagccagaag tttgagccca ccggcgtcgg ccacctggag 600
accctccaca tggccatgtc ctggcaggac cacggccgga tcctgcgctg ccagctctcc 660
gtggccaatc acagggctca gagcgagatt cacctccaag tgaagtatgc ccccaagggt 720
gtgaagatcc tcctcagccc ctcggggagg aacatccttc caggtgagct ggtcacactc 780
acctgccagg tgaacagcag ctaccctgca gtcagttcca ttaagtggct caaggatggg 840
gtacgcctcc aaaccaagac tggtgtgctg cacctgcccc aggcagcctg gagcgatgct 900
ggcgtctaca cctgccaagc tgagaacggc gtgggctctt tggtctcacc ccccatcagc 960
ctccacatct tcatggctga ggtccaggtg agcccagcag gtcccatcct ggagaaccag 1020
acagtgacac tagtctgcaa cacacccaat gaggcaccca gtgatctccg ctacagctgg 1080
tacaagaacc atgtcctgct ggaggatgcc cactcccata ccctccggct gcacttggcc 1140
actagggctg atactggctt ctacttctgt gaggtgcaga acgtccatgg cagcgagcgc 1200
tcgggccctg tcagcgtggt agtcaaccac ccgcctctca ctccagtcct gacagccttc 1260
ctggagaccc aggcgggact tgtgggcatc cttcactgct ctgtggtcag tgagcccctg 1320
gccacactgg tgctgtcaca tgggggtcat atcctggcct ccacctccgg ggacagtgat 1380
cacagcccac gcttcagtgg tacctctggt cccaactccc tgcgcctgga gatccgagac 1440
ctggaggaaa ctgacagtgg ggagtacaag tgctcagcca ccaactccct tggaaatgca 1500
acctccaccc tggacttcca tgccaatgcc gcccgtctcc tcatcagccc ggcagccgag 1560
gtggtggaag gacaggcagt gacactgagc tgcagaagcg gcctaagccc cacacctgat 1620
gcccgcttct cctggtacct gaatggagcc ctgcttcacg agggtcccgg cagcagcctc 1680
ctgctccccg cggcctccag cactgacgcc ggctcatacc actgccgggc ccgggacggc 1740
cacagtgcca gtggcccctc ttcgccagct gttctcactg tgctctaccc ccctcgacaa 1800
ccaacattca ccaccaggct ggaccttgat gccgctgggg ccggggctgg acggcgaggc 1860
ctccttttgt gccgtgtgga cagcgacccc cccgccaggc tgcagctgct ccacaaggac 1920
cgtgttgtgg ccacttccct gccatcaggg ggtggctgca gcacctgtgg gggctgttcc 1980
ccacgcatga aggtcaccaa agcccccaac ttgctgcgtg tggagattca caaccctttg 2040
ctggaagagg agggcttgta cctctgtgag gccagcaatg ccctgggcaa cgcctccacc 2100
tcagccacct tcaatggcca ggccactgtc ctggccattg caccatcaca cacacttcag 2160
gagggcacag aagccaactt gacttgcaac gtgagccggg aagctgctgg cagccctgct 2220
aacttctcct ggttccgaaa tggggtgctg tgggcccagg gtcccctgga gaccgtgaca 2280
ctgctgcccg tggccagaac tgatgctgcc ctttacgcct gccgcatcct gactgaggct 2340
ggtgcccagc tctccactcc cgtgctcctg agtgtactct atcccccgga ccgtccaaag 2400
ctgtcagccc tcctagacat gggccagggc cacatggctc tgttcatctg cactgtggac 2460
agccgccccc tggccttgct ggccttgttc catggggagc acctcctggc caccagcctg 2520
ggtccccagg tcccatccca tggtcggttc caggctaaag ctgaggccaa ctccctgaag 2580
ttagaggtcc gagaactggg ccttggggac tctggcagct accgctgtga ggccacaaat 2640
gttcttggat catccaacac ctcactcttc ttccaggtcc gaggagcctg ggtccaggtg 2700
tcaccatcac ctgagctcca agagggccag gctgtggtcc tgagctgcca ggtacacaca 2760
ggagtcccag aggggacctc atatcgttgg tatcgggatg gccagcccct ccaggagtcg 2820
acctcggcca cgctccgctt tgcagccata actttgacac aagctggggc ctatcattgc 2880
caagcccagg ccccaggctc agccaccacg agcctagctg cacccatcag cctccacgtg 2940
tcctatgccc cacgccacgt cacactcact accctgatgg acacaggccc tggacgactg 3000
ggcctcctcc tgtgccgtgt ggacagtgac cctccggccc agctgcggct gctccacggg 3060
gatcgccttg tggcctccac cctacaaggt gtggggggac ccgaaggcag ctctcccagg 3120
ctgcatgtgg ctgtggcccc caacacactg cgtctggaga tccacggggc tatgctggag 3180
gatgagggtg tctatatctg tgaggcctcc aacaccctgg gccaggcctc ggcctcagct 3240
gacttcgacg ctcaagctgt gaatgtgcag gtgtggcccg gggctaccgt gcgggagggg 3300
cagctggtga acctgacctg ccttgtgtgg accactcacc cggcccagct cacctacaca 3360
tggtaccagg atgggcagca gcgcctggat gcccactcca tccccctgcc caacgtcaca 3420
gtcagggatg ccacctccta ccgctgcggt gtgggccccc ctggtcgggc accccgcctc 3480
tccagaccta tcaccttgga cgtcctctac gcgccccgca acctgcgcct gacctacctc 3540
ctggagagcc atggcgggca gctggccctg gtactgtgca ctgtggacag ccgcccgccc 3600
gcccagctgg ccctcagcca cgccggtcgc ctcttggcct cctcgacagc agcctctgtc 3660
cccaacaccc tgcgcctgga gctgcgaggg ccacagccca gggatgaggg tttctacagc 3720
tgctctgccc gcagccctct gggccaggcc aacacgtccc tggagctgcg gctggagggt 3780
gtgcgggtga tcctggctcc ggaggctgcc gtgcctgaag gtgcccccat cacagtgacc 3840
tgtgcggacc ctgctgccca cgcacccaca ctctatactt ggtaccacaa cggtcgttgg 3900
ctgcaggagg gtccagctgc ctcactctca ttcctggtgg ccacgcgggc tcatgcaggc 3960
gcctactctt gccaggccca ggatgcccag ggcacccgca gctcccgtcc tgctgccctg 4020
caagtcctct atgcccctca ggacgctgtc ctgtcctcct tccgggactc cagggccaga 4080
tccatggctg tgatacagtg cactgtggac agtgagccac ctgctgagct ggccctatct 4140
catgatggca aggtgctggc cacgagcagc ggggtccaca gcttggcatc agggacaggc 4200
catgtccagg tggcccgaaa cgccctacgg ctgcaggtgc aagatgtgcc tgcaggtgat 4260
gacacctatg tttgcacagc ccaaaacttg ctgggctcaa tcagcaccat cgggcggttg 4320
caggtagaag gtgcacgcgt ggtggcagag cctggcctgg acgtgcctga gggcgctgcc 4380
ctgaacctca gctgccgcct cctgggtggc cctgggcctg tgggcaactc cacctttgca 4440
tggttctgga atgaccggcg gctgcacgcg gagcctgtgc ccactctcgc cttcacccac 4500
gtggctcgtg ctcaagctgg gatgtaccac tgcctggctg agctccccac tggggctgct 4560
gcctctgctc cagtcatgct ccgtgtgctc taccctccca agacgcccac catgatggtc 4620
ttcgtggagc ctgagggtgg cctccggggc atcctggatt gccgagtgga cagcgagccg 4680
ctcgccagcc tgactctcca ccttggcagt cgactggtgg cctccagtca gccccagggt 4740
gctcctgcag agccacacat ccatgtcctg gcttccccca atgccctgag ggtggacatc 4800
gaggcgctga ggcccagcga ccaaggggaa tacatctgtt ctgcctcaaa tgtcctgggc 4860
tctgcctcta cctccaccta ctttggggtc agagccctgc accgcctgca tcagttccag 4920
cagctgctct gggtcctggg actgctggtg ggcctcctgc tcctgctgtt gggcctgggg 4980
gcctgctaca cctggagaag gaggcgtgtt tgtaagcaga gcatgggcga gaattcggtg 5040
gagatggctt ttcagaaaga gaccacgcag ctcattgatc ctgatgcagc cacatgtgag 5100
acctcaacct gtgccccacc cctgggctga ccagtggtgt tgcctgccct ccggaggaga 5160
aagtggccag aatctgtgat gactccagcc tatgaatgtg aatgaggcag tgttgagtcc 5220
tgcccgcctc tacgaaaaca gctctgtgac atctgacttt ttatgacctg gccccaagcc 5280
tcttgccccc ccaaaaatgg gtggtgagag gtctgcccag gagggtgttg accctggagg 5340
acactgaaga gcactgagct gatctcgctc tctcttctct ggatctcctc ccttctctcc 5400
atttctccct caaaggaagc cctgcccttt cacatccttc tcctcgaaag tcaccctgga 5460
ctttggttgg attgcagcat cctgcatcct cagaggctca ccaaggcatt ctgtattcaa 5520
cagagtatca gtcagcctgc tctaacaaga gaccaaatac agtgacttca acatgataga 5580
attttatttt tctctcccac gctagtctgg ctgttacgat ggtttatgat gttggggctc 5640
aggatccttc tatcttcctt ttctctatcc ctaaaatgat gcctttgatt gtgaggctca 5700
ccatggcccc gctttgtcca catgccctcc agccagaaga aggaagagtg gaggtagaag 5760
cacacccatg cccatggtgg acgcaactca gaagctgcac aggacttttc cactcacttc 5820
ccattggctg gagtattgtc acatggctac tgcaagctac aagggagact gggaaatgta 5880
gtttttattt tgagtccaga ggacatttgg aattggactt ccaaaggact cccaactgtg 5940
agctcatccc tgagactttt gacattgttg ggaatgccac cagcaggcca tgttttgtct 6000
cagtgcccat ctactgaggg ccagggtgtg cccctggcca ttctggttgt gggcttcctg 6060
gaagaggtga tcactctcac actaagactg aggaaataaa aaaggtttgg tgttttccta 6120
gggagagagc atgccaggca gtggagttgc ctaagcagac atccttgtgc cagatttggc 6180
ccctgaaaga agagatgccc tcattcccac caccaccccc cctaccccca gggactgggt 6240
actaccttac tggcccttac aagagtggag ggcagacaca gatgttgtca gcatccttat 6300
tcctgctcca gatgcatctc tgttcatgac tgtgtgagct cctgtccttt tcctggagac 6360
cctgtgtcgg gctgttaaag agaatgagtt accaagaagg aatgacgtgc ccctgcgaat 6420
cagggaccaa caggagagag ctcttgagtg ggctagtgac tccccctgca gcctggtgga 6480
gatggtgtga ggagcgaaga gccctctgct ctaggatttg ggttgaaaaa cagagagaga 6540
agtggggagt tgccacagga gctaacacgc tgggaggcag ttgggggcgg gtgaactttg 6600
tgtagccgag gccgcaccct ccctcattcc aggctcattc attttcatgc tccattgcca 6660
gactcttgct gggagcccgt ccagaatgtc ctcccaataa aactccatcc tatgacgcaa 6720
aaaaaaaaaa aaaaaa 6736
<210> SEQ ID NO 14
<211> LENGTH: 1709
<212> TYPE: PRT
<213> ORGANISM: homo sapiens
<300> PUBLICATION INFORMATION:
<308> DATABASE ACCESSION NUMBER: GenPept/NP_075556
<309> DATABASE ENTRY DATE: 2001-02-13
<313> RELEVANT RESIDUES IN SEQ ID NO: (1)..(1709)
<400> SEQUENCE: 14
Met Gly Phe Leu Pro Lys Leu Leu Leu Leu Ala Ser Phe Phe Pro Ala
1 5 10 15
Gly Gln Ala Ser Trp Gly Val Ser Ser Pro Gln Asp Val Gln Gly Val
20 25 30
Lys Gly Ser Cys Leu Leu Ile Pro Cys Ile Phe Ser Phe Pro Ala Asp
35 40 45
Val Glu Val Pro Asp Gly Ile Thr Ala Ile Trp Tyr Tyr Asp Tyr Ser
50 55 60
Gly Gln Arg Gln Val Val Ser His Ser Ala Asp Pro Lys Leu Val Glu
65 70 75 80
Ala Arg Phe Arg Gly Arg Thr Glu Phe Met Gly Asn Pro Glu His Arg
85 90 95
Val Cys Asn Leu Leu Leu Lys Asp Leu Gln Pro Glu Asp Ser Gly Ser
100 105 110
Tyr Asn Phe Arg Phe Glu Ile Ser Glu Val Asn Arg Trp Ser Asp Val
115 120 125
Lys Gly Thr Leu Val Thr Val Thr Glu Glu Pro Arg Val Pro Thr Ile
130 135 140
Ala Ser Pro Val Glu Leu Leu Glu Gly Thr Glu Val Asp Phe Asn Cys
145 150 155 160
Ser Thr Pro Tyr Val Cys Leu Gln Glu Gln Val Arg Leu Gln Trp Gln
165 170 175
Gly Gln Asp Pro Ala Arg Ser Val Thr Phe Asn Ser Gln Lys Phe Glu
180 185 190
Pro Thr Gly Val Gly His Leu Glu Thr Leu His Met Ala Met Ser Trp
195 200 205
Gln Asp His Gly Arg Ile Leu Arg Cys Gln Leu Ser Val Ala Asn His
210 215 220
Arg Ala Gln Ser Glu Ile His Leu Gln Val Lys Tyr Ala Pro Lys Gly
225 230 235 240
Val Lys Ile Leu Leu Ser Pro Ser Gly Arg Asn Ile Leu Pro Gly Glu
245 250 255
Leu Val Thr Leu Thr Cys Gln Val Asn Ser Ser Tyr Pro Ala Val Ser
260 265 270
Ser Ile Lys Trp Leu Lys Asp Gly Val Arg Leu Gln Thr Lys Thr Gly
275 280 285
Val Leu His Leu Pro Gln Ala Ala Trp Ser Asp Ala Gly Val Tyr Thr
290 295 300
Cys Gln Ala Glu Asn Gly Val Gly Ser Leu Val Ser Pro Pro Ile Ser
305 310 315 320
Leu His Ile Phe Met Ala Glu Val Gln Val Ser Pro Ala Gly Pro Ile
325 330 335
Leu Glu Asn Gln Thr Val Thr Leu Val Cys Asn Thr Pro Asn Glu Ala
340 345 350
Pro Ser Asp Leu Arg Tyr Ser Trp Tyr Lys Asn His Val Leu Leu Glu
355 360 365
Asp Ala His Ser His Thr Leu Arg Leu His Leu Ala Thr Arg Ala Asp
370 375 380
Thr Gly Phe Tyr Phe Cys Glu Val Gln Asn Val His Gly Ser Glu Arg
385 390 395 400
Ser Gly Pro Val Ser Val Val Val Asn His Pro Pro Leu Thr Pro Val
405 410 415
Leu Thr Ala Phe Leu Glu Thr Gln Ala Gly Leu Val Gly Ile Leu His
420 425 430
Cys Ser Val Val Ser Glu Pro Leu Ala Thr Leu Val Leu Ser His Gly
435 440 445
Gly His Ile Leu Ala Ser Thr Ser Gly Asp Ser Asp His Ser Pro Arg
450 455 460
Phe Ser Gly Thr Ser Gly Pro Asn Ser Leu Arg Leu Glu Ile Arg Asp
465 470 475 480
Leu Glu Glu Thr Asp Ser Gly Glu Tyr Lys Cys Ser Ala Thr Asn Ser
485 490 495
Leu Gly Asn Ala Thr Ser Thr Leu Asp Phe His Ala Asn Ala Ala Arg
500 505 510
Leu Leu Ile Ser Pro Ala Ala Glu Val Val Glu Gly Gln Ala Val Thr
515 520 525
Leu Ser Cys Arg Ser Gly Leu Ser Pro Thr Pro Asp Ala Arg Phe Ser
530 535 540
Trp Tyr Leu Asn Gly Ala Leu Leu His Glu Gly Pro Gly Ser Ser Leu
545 550 555 560
Leu Leu Pro Ala Ala Ser Ser Thr Asp Ala Gly Ser Tyr His Cys Arg
565 570 575
Ala Arg Asp Gly His Ser Ala Ser Gly Pro Ser Ser Pro Ala Val Leu
580 585 590
Thr Val Leu Tyr Pro Pro Arg Gln Pro Thr Phe Thr Thr Arg Leu Asp
595 600 605
Leu Asp Ala Ala Gly Ala Gly Ala Gly Arg Arg Gly Leu Leu Leu Cys
610 615 620
Arg Val Asp Ser Asp Pro Pro Ala Arg Leu Gln Leu Leu His Lys Asp
625 630 635 640
Arg Val Val Ala Thr Ser Leu Pro Ser Gly Gly Gly Cys Ser Thr Cys
645 650 655
Gly Gly Cys Ser Pro Arg Met Lys Val Thr Lys Ala Pro Asn Leu Leu
660 665 670
Arg Val Glu Ile His Asn Pro Leu Leu Glu Glu Glu Gly Leu Tyr Leu
675 680 685
Cys Glu Ala Ser Asn Ala Leu Gly Asn Ala Ser Thr Ser Ala Thr Phe
690 695 700
Asn Gly Gln Ala Thr Val Leu Ala Ile Ala Pro Ser His Thr Leu Gln
705 710 715 720
Glu Gly Thr Glu Ala Asn Leu Thr Cys Asn Val Ser Arg Glu Ala Ala
725 730 735
Gly Ser Pro Ala Asn Phe Ser Trp Phe Arg Asn Gly Val Leu Trp Ala
740 745 750
Gln Gly Pro Leu Glu Thr Val Thr Leu Leu Pro Val Ala Arg Thr Asp
755 760 765
Ala Ala Leu Tyr Ala Cys Arg Ile Leu Thr Glu Ala Gly Ala Gln Leu
770 775 780
Ser Thr Pro Val Leu Leu Ser Val Leu Tyr Pro Pro Asp Arg Pro Lys
785 790 795 800
Leu Ser Ala Leu Leu Asp Met Gly Gln Gly His Met Ala Leu Phe Ile
805 810 815
Cys Thr Val Asp Ser Arg Pro Leu Ala Leu Leu Ala Leu Phe His Gly
820 825 830
Glu His Leu Leu Ala Thr Ser Leu Gly Pro Gln Val Pro Ser His Gly
835 840 845
Arg Phe Gln Ala Lys Ala Glu Ala Asn Ser Leu Lys Leu Glu Val Arg
850 855 860
Glu Leu Gly Leu Gly Asp Ser Gly Ser Tyr Arg Cys Glu Ala Thr Asn
865 870 875 880
Val Leu Gly Ser Ser Asn Thr Ser Leu Phe Phe Gln Val Arg Gly Ala
885 890 895
Trp Val Gln Val Ser Pro Ser Pro Glu Leu Gln Glu Gly Gln Ala Val
900 905 910
Val Leu Ser Cys Gln Val His Thr Gly Val Pro Glu Gly Thr Ser Tyr
915 920 925
Arg Trp Tyr Arg Asp Gly Gln Pro Leu Gln Glu Ser Thr Ser Ala Thr
930 935 940
Leu Arg Phe Ala Ala Ile Thr Leu Thr Gln Ala Gly Ala Tyr His Cys
945 950 955 960
Gln Ala Gln Ala Pro Gly Ser Ala Thr Thr Ser Leu Ala Ala Pro Ile
965 970 975
Ser Leu His Val Ser Tyr Ala Pro Arg His Val Thr Leu Thr Thr Leu
980 985 990
Met Asp Thr Gly Pro Gly Arg Leu Gly Leu Leu Leu Cys Arg Val Asp
995 1000 1005
Ser Asp Pro Pro Ala Gln Leu Arg Leu Leu His Gly Asp Arg Leu
1010 1015 1020
Val Ala Ser Thr Leu Gln Gly Val Gly Gly Pro Glu Gly Ser Ser
1025 1030 1035
Pro Arg Leu His Val Ala Val Ala Pro Asn Thr Leu Arg Leu Glu
1040 1045 1050
Ile His Gly Ala Met Leu Glu Asp Glu Gly Val Tyr Ile Cys Glu
1055 1060 1065
Ala Ser Asn Thr Leu Gly Gln Ala Ser Ala Ser Ala Asp Phe Asp
1070 1075 1080
Ala Gln Ala Val Asn Val Gln Val Trp Pro Gly Ala Thr Val Arg
1085 1090 1095
Glu Gly Gln Leu Val Asn Leu Thr Cys Leu Val Trp Thr Thr His
1100 1105 1110
Pro Ala Gln Leu Thr Tyr Thr Trp Tyr Gln Asp Gly Gln Gln Arg
1115 1120 1125
Leu Asp Ala His Ser Ile Pro Leu Pro Asn Val Thr Val Arg Asp
1130 1135 1140
Ala Thr Ser Tyr Arg Cys Gly Val Gly Pro Pro Gly Arg Ala Pro
1145 1150 1155
Arg Leu Ser Arg Pro Ile Thr Leu Asp Val Leu Tyr Ala Pro Arg
1160 1165 1170
Asn Leu Arg Leu Thr Tyr Leu Leu Glu Ser His Gly Gly Gln Leu
1175 1180 1185
Ala Leu Val Leu Cys Thr Val Asp Ser Arg Pro Pro Ala Gln Leu
1190 1195 1200
Ala Leu Ser His Ala Gly Arg Leu Leu Ala Ser Ser Thr Ala Ala
1205 1210 1215
Ser Val Pro Asn Thr Leu Arg Leu Glu Leu Arg Gly Pro Gln Pro
1220 1225 1230
Arg Asp Glu Gly Phe Tyr Ser Cys Ser Ala Arg Ser Pro Leu Gly
1235 1240 1245
Gln Ala Asn Thr Ser Leu Glu Leu Arg Leu Glu Gly Val Arg Val
1250 1255 1260
Ile Leu Ala Pro Glu Ala Ala Val Pro Glu Gly Ala Pro Ile Thr
1265 1270 1275
Val Thr Cys Ala Asp Pro Ala Ala His Ala Pro Thr Leu Tyr Thr
1280 1285 1290
Trp Tyr His Asn Gly Arg Trp Leu Gln Glu Gly Pro Ala Ala Ser
1295 1300 1305
Leu Ser Phe Leu Val Ala Thr Arg Ala His Ala Gly Ala Tyr Ser
1310 1315 1320
Cys Gln Ala Gln Asp Ala Gln Gly Thr Arg Ser Ser Arg Pro Ala
1325 1330 1335
Ala Leu Gln Val Leu Tyr Ala Pro Gln Asp Ala Val Leu Ser Ser
1340 1345 1350
Phe Arg Asp Ser Arg Ala Arg Ser Met Ala Val Ile Gln Cys Thr
1355 1360 1365
Val Asp Ser Glu Pro Pro Ala Glu Leu Ala Leu Ser His Asp Gly
1370 1375 1380
Lys Val Leu Ala Thr Ser Ser Gly Val His Ser Leu Ala Ser Gly
1385 1390 1395
Thr Gly His Val Gln Val Ala Arg Asn Ala Leu Arg Leu Gln Val
1400 1405 1410
Gln Asp Val Pro Ala Gly Asp Asp Thr Tyr Val Cys Thr Ala Gln
1415 1420 1425
Asn Leu Leu Gly Ser Ile Ser Thr Ile Gly Arg Leu Gln Val Glu
1430 1435 1440
Gly Ala Arg Val Val Ala Glu Pro Gly Leu Asp Val Pro Glu Gly
1445 1450 1455
Ala Ala Leu Asn Leu Ser Cys Arg Leu Leu Gly Gly Pro Gly Pro
1460 1465 1470
Val Gly Asn Ser Thr Phe Ala Trp Phe Trp Asn Asp Arg Arg Leu
1475 1480 1485
His Ala Glu Pro Val Pro Thr Leu Ala Phe Thr His Val Ala Arg
1490 1495 1500
Ala Gln Ala Gly Met Tyr His Cys Leu Ala Glu Leu Pro Thr Gly
1505 1510 1515
Ala Ala Ala Ser Ala Pro Val Met Leu Arg Val Leu Tyr Pro Pro
1520 1525 1530
Lys Thr Pro Thr Met Met Val Phe Val Glu Pro Glu Gly Gly Leu
1535 1540 1545
Arg Gly Ile Leu Asp Cys Arg Val Asp Ser Glu Pro Leu Ala Ser
1550 1555 1560
Leu Thr Leu His Leu Gly Ser Arg Leu Val Ala Ser Ser Gln Pro
1565 1570 1575
Gln Gly Ala Pro Ala Glu Pro His Ile His Val Leu Ala Ser Pro
1580 1585 1590
Asn Ala Leu Arg Val Asp Ile Glu Ala Leu Arg Pro Ser Asp Gln
1595 1600 1605
Gly Glu Tyr Ile Cys Ser Ala Ser Asn Val Leu Gly Ser Ala Ser
1610 1615 1620
Thr Ser Thr Tyr Phe Gly Val Arg Ala Leu His Arg Leu His Gln
1625 1630 1635
Phe Gln Gln Leu Leu Trp Val Leu Gly Leu Leu Val Gly Leu Leu
1640 1645 1650
Leu Leu Leu Leu Gly Leu Gly Ala Cys Tyr Thr Trp Arg Arg Arg
1655 1660 1665
Arg Val Cys Lys Gln Ser Met Gly Glu Asn Ser Val Glu Met Ala
1670 1675 1680
Phe Gln Lys Glu Thr Thr Gln Leu Ile Asp Pro Asp Ala Ala Thr
1685 1690 1695
Cys Glu Thr Ser Thr Cys Ala Pro Pro Leu Gly
1700 1705
<210> SEQ ID NO 15
<211> LENGTH: 40
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: PCR forward primer CD163
<400> SEQUENCE: 15
cac cat gga caa act cag aat ggt gct aca tga aaa ctc t 40
<210> SEQ ID NO 16
<211> LENGTH: 33
<212> TYPE: DNA
<213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: PCR reverse primer CD163
<400> SEQUENCE: 16
tca ttg tac ttc aga gtg gtc tcc tga ggg att 33
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