Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification
Reexamination Certificate
2005-03-15
2005-03-15
Prouty, Rebecca E. (Department: 1652)
Chemistry: molecular biology and microbiology
Process of mutation, cell fusion, or genetic modification
C436S086000, C436S002000, C435S029000, C435S069100, C435S252300, C435S325000, C435S320100, C536S023400
Reexamination Certificate
active
06867042
ABSTRACT:
A solubility reporter for measuring a protein's solubility in vivo or in vitro is described. The reporter, which can be used in a single living cell, gives a specific signal suitable for determining whether the cell bears a soluble version of the protein of interest. A pool of random mutants of an arbitrary protein, generated using error-prone in vitro recombination, may also be screened for more soluble versions using the reporter, and these versions may be recombined to yield variants having further-enhanced solubility. The method of the present invention includes “irrational” (random mutagenesis) methods, which do not require a priori knowledge of the three-dimensional structure of the protein of interest. Multiple sequences of mutation/genetic recombination and selection for improved solubility are demonstrated to yield versions of the protein which display enhanced solubility.
REFERENCES:
patent: 5491084 (1996-02-01), Chalfie et al.
Willem P.C. Stemmer, “Rapid Evolution Of A Protein In Vitro By DNA Shuffling,” Nature 370, 389 (1994).
Willem P.C. Stemmer, “DNA Shuffling By Random Fragmentation And Reassembly: In Vitro Recombination For Molecular Evolution,” Proc. Natl. Acad. Sci. USA 91, 10747 (1994).
Willem P.C. Stemmer, “Searching Sequence Space: Using Recombination to Search More Efficiently and Thoroughly Instead of Making Bigger Combinatorial Libraries,” Biotechnology 13, 549 (1995).
Frances H. Arnold, “Directed Evolution: Creating Biocatalysts for the Future,” Chemical Engineering Science 51, 5091 (1996).
Ji-Hu Zhang et al., “Directed Evolution Of A Fucosidase from a Galactosidase by DNA Shuffling and Screening,” Proc. Natl. Acad. Sci. USA 94, 4504 (1997).
Huimin Zhao et al., “Functional and Nonfunctional Mutations Distinguished by Random Recombination of Homologous Genes,” Proc. Natl. Acad. Sci. USA 94, 99 (1997).
Jeffrey C. Moore et al., “Strategies for the In Vitro Evolution of Protein Function: Enzyme Evolution by Random Recombination of Improved Sequences”, J. Mol. Biol. 272, 336 (1997).
Andreas Crameri et al., “Improved Green Fluorescent Protein by Molecular Evolution Using DNA Shuffling,” Nature Biotechnology 14, 315 (1996).
Roger Heim et al., “Wavelength Mutations and Posttranslational Autoxidation of Green Fluorescent Protein,” Proc. Natl. Acad. Sci. USA 91, 12501 (1994).
Chun Wu et al., “Novel Green Fluorescent Protein (GFP) Baculovirus Expression Vectors,” 190, 157 (1997).
N. Garamszegi et al., “Application of a Chimeric Green Fluorescent Protein to Study Protein-Protein Interactions,” Biotechniques 23, 864 (1997).
G. MacBeath, et al., “Redesigning Enzyme Topology by Directed Evolution,” Science 279, 1958 (1998).
Pierre Martineau et al., “Expression of an Antibody Fragment at High Levels in the Bacterial Cytoplasm,” J. Mol. Biol. 280, 117 (1998).
Karl Proba et al., “Antibody scFv Fragments Without Disulfide Bonds Made by Molecular Evolution,” J. Mol. Biol. 275, 245 (1998).
Zhanglin Lin et al., “Functional Expression of Horseradish Peroxidase inE. coliby Directed Evolution,” Biotechnol. Prog. 15, 467 (1999).
Geoffrey S. Waldo et al., “Rapid Protein-Folding Assay Using Green Fluorescent Protein” Nature Biotechnology 17, 691 (1999).
Russell Higuchi, “Recombinant PCR” in PCR Protocols, a Guide to Methods and Applications , Michael A. Innis, David H. Gelfand, John J. Sninsky, and Thomas J. White, eds. Academic press, Inc., 177 (1990).
Roger Helm et al., “Improved Green Fluorescence,” Nature 373, 663 (1995).
Sorel Fitz-Gibbon et al., “A Fosmid-based Genomic Map and Identification of 474 Genes of the Hyperthermophilic ArchaeonPyrobaculum aerophilum.” Extremophiles 1, 36 (1997).
Geoffrey Zubay, “In Vitro Synthesis of Protein in Microbial Systems.” Ann. Rev. Genet. 7, 267 (1973).
Frederick C. Neidhardt, “Chemical Composition ofEscherichia coli,” Neidhardt, F. C., ed. inEscherichia coliandSalmonella typhimurium: Cellular and Molecular Biology, pp. 3-6, American Society of Microbiology, Washington, D. C. (1987).
Yang Zhang et al., “Expression of Eukaryotic Proteins in Soluble Form inEscherichia coli”, Protein Expr. Purif. 12, 159 (1998).
Humin Zhao et al., “Optimization of DNA Shuffling for High Fidelity Recombination.” Nuc. Acids Res. 25, 1307 (1997).
Thomas C. Terwilliger et al., “In-vivo Characterization of Mutants of the Bacteriophage-f1 Gene V Protein Isolated by Saturation Mutagenesis.” J. Mol. Biol. 236, 556 (1994).
Lynn F. Dickey et al., “Differences in the Regulation of Messenger-RNA for Housekeeping and Specialized-Cell Ferritin: A Comparison of Three Distinct Ferritin Complementary DNAs, the Corresponding Subunits, and Identification of the First Processed Pseudogene in Amphibia.” J. Biol. Chem. 262, 7901 (1987).
Geoffrey S. Waldo et al., “Ferritin and Iron Biomineralization”, Comprehensive Supramolecular Chemistry 5, pp. 65-91, Susslick, K. vol. ed., Pergamon Press, U.K., (1996).
P. M. Harrison and P. Arosio, “The Ferritins: Molecular Properties, Iron Storage Function and Cellular Regulation.” Biochim. et Biophys. Acta. 1275, 161 (1996).
T. Moors et al., “A Sensitive Post-DAB Enhancement Technique for Demonstration of Iron in the Central-Nervous System.” Histochem. 99, 471 (1993).
Malcolm J. Casadaban et al., “β-Galactosidase Gene Fusions for Analyzing Gene Expression in Escherichia Coli and Yeast,” Methods Enzymol. 100, 293 (1983).
Maria T. Martin et al., “Determination of Polyprotein Processing Sites by Amino Terminal Sequencing of Nonstructural Proteins Encoded by Plum Pox Polyvirus”, Virus Res. 15, 97 (1990).
Rolf Lutz et al., “Independent and Tight Regulation of Transcriptional Units inE. colivia the LacR/O, the TetR/O and AraD/l1-l2Regulatory Elements”, Nucleic Acids Res., 25(6), 1203 (1997).
Freund Samuel M.
Prouty Rebecca E.
Ramirez Delia M.
Sharples Kenneth K.
The Regents of the University of California
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