Patent application number | Description | Published |
20090068732 | Directed evolution methods for improving polypeptide folding and solubility and superfolder fluorescent proteins generated thereby - The current invention provides methods of improving folding of polypeptides using a poorly folding domain as a component of a fusion protein comprising the poorly folding domain and a polypeptide of interest to be improved. The invention also provides novel green fluorescent proteins (GFPs) and red fluorescent proteins that have enhanced folding properties. | 03-12-2009 |
20090142820 | Directed evolution methods for improving polypeptide folding, solubility and stability - The invention provides directed evolution methods for improving the folding, solubility and stability (including thermostability) characteristics of polypeptides. In one aspect, the invention provides a method for generating folding and stability-enhanced variants of proteins, including but not limited to fluorescent proteins, chromophoric proteins and enzymes. In another aspect, the invention provides methods for generating thermostable variants of a target protein or polypeptide via an internal destabilization baiting strategy. Internally destabilization a protein of interest is achieved by inserting a heterologous, folding-destabilizing sequence (folding interference domain) within DNA encoding the protein of interest, evolving the protein sequences adjacent to the heterologous insertion to overcome the destabilization (using any number of mutagenesis methods), thereby creating a library of variants. The variants in the library are expressed, and those with enhanced folding characteristics selected. | 06-04-2009 |
20090170721 | Circular permutant GFP insertion folding reporters - Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited. | 07-02-2009 |
20100222551 | Highly thermostable fluorescent proteins - Thermostable fluorescent proteins (TSFPs), methods for generating these and other stability-enhanced proteins, polynucleotides encoding such proteins, and assays and method for using the TSFPs and TSFP-encoding nucleic acid molecules are provided. The TSFPs of the invention show extremely enhanced levels of stability and thermotolerance. In one case, for example, a TSFP of the invention is so stable it can be heated to 99° C. for short periods of time without denaturing, and retains 85% of its fluorescence when heated to 80° C. for several minutes. The invention also provides a method for generating stability-enhanced variants of a protein, including but not limited to fluorescent proteins. | 09-02-2010 |
20100261267 | Circular permutant GFP insertion folding reporters - Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited. | 10-14-2010 |
20110245464 | HIGHLY THERMOSTABLE FLUORESCENT PROTEINS - Thermostable fluorescent proteins (TSFPs), methods for generating these and other stability-enhanced proteins, polynucleotides encoding such proteins, and assays and method for using the TSFPs and TSFP-encoding nucleic acid molecules are provided. The TSFPs of the invention show extremely enhanced levels of stability and thermotolerance. In one case, for example, a TSFP of the invention is so stable it can be heated to 99° C. for short periods of time without denaturing, and retains 85% of its fluorescence when heated to 80° C. for several minutes. The invention also provides a method for generating stability-enhanced variants of a protein, including but not limited to fluorescent proteins. | 10-06-2011 |
20110256620 | NUCLEIC ACID ENCODING A SELF-ASSEMBLING SPLIT-FLUORESCENT PROTEIN SYSTEM - The invention provides a protein labeling and detection system based on self-complementing fragments of fluorescent and chromophoric proteins. The system of the invention is exemplified with various combinations of self-complementing fragments derived from | 10-20-2011 |
20120077266 | HIGHLY THERMOSTABLE FLUORESCENT PROTEINS - Thermostable fluorescent proteins (TSFPs), methods for generating these and other stability-enhanced proteins, polynucleotides encoding such proteins, and assays and method for using the TSFPs and TSFP-encoding nucleic acid molecules are provided. The TSFPs of the invention show extremely enhanced levels of stability and thermotolerance. In one case, for example, a TSFP of the invention is so stable it can be heated to 99° C. for short periods of time without denaturing, and retains 85% of its fluorescence when heated to 80° C. for several minutes. The invention also provides a method for generating stability-enhanced variants of a protein, including but not limited to fluorescent proteins. | 03-29-2012 |
20120282643 | CYAN AND YELLOW FLUORESCENT COLOR VARIANTS OF SPLIT GFP - Disclosed herein are Split-Fluorescent proteins (SFPs) including Split-Yellow Fluorescent Proteins and Split-Cyan Fluorescent proteins. Further disclosed are methods of using SFPs. For example, methods of identifying the subcellular localization of a protein and methods of identifying the membrane topology of a membrane protein are disclosed herein. | 11-08-2012 |
20120329673 | VECTORS AND METHODS FOR SELECTING OPEN READING FRAMES - The invention provides vectors and methods designed for screening large random DNA fragment libraries for the presence of open reading frames that are free of both internal ribosome binding sites (IRBS) and stop codons. The invention overcomes the principal limitation of known ORF-selector systems, namely the potential for ORF-induced folding interference of the downstream fused reporter, by not fusing the reporter to the ORF, but rather by providing a mechanism for coupled translation of an unfused downstream reporter. | 12-27-2012 |
20130040384 | CIRCULAR PERMUTANT GFP INSERTION FOLDING REPORTERS - Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited. | 02-14-2013 |
20140024555 | METHOD OF IDENTIFYING SOLUBLE PROTEINS AND SOLUBLE PROTEIN COMPLEXES - Provided herein are methods of identifying a protein as soluble, as well as methods of identifying a soluble protein complex of at least two proteins. The methods allow for the determination of in vitro solubility, or both in vitro and in vivo solubility, of a protein or protein complex. | 01-23-2014 |
20140141415 | NUCLEIC ACID ENCODING A SELF-ASSEMBLING SPLIT-FLUORESCENT PROTEIN SYSTEM - The invention provides a protein labeling and detection system based on self-complementing fragments of fluorescent and chromophoric proteins. The system of the invention is exemplified with various combinations of self-complementing fragments derived from | 05-22-2014 |