Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues
Reexamination Certificate
2007-08-21
2007-08-21
Monshipouri, Maryam (Department: 1656)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
C435S069100, C435S068100, C435S069700
Reexamination Certificate
active
11329401
ABSTRACT:
The current invention provides methods for producing a polypeptide as inclusion bodies in bacterial host cells. The present methods are carried out by forming a gene construct comprising the genetic sequence encoding a polypeptide operatively linked to that of an inclusion partner protein, such asE. colithioredoxin or a modifiedE. colithioredoxin, such that host cells comprising the gene construct produce the polypeptide as intracellular inclusion bodies. The methods of the present invention facilitate the rapid isolation and purification of recombinant proteins. In addition, the present methods may be useful for producing polypeptides or proteins which are small and are typically difficult to express, as well as those proteins that are toxic to host cells such asE. coli. The present invention also provides plasmids, vectors and host cells to be used in the present invention for production of polypeptides, and methods of production of polypeptides using these vectors and host cells. The invention further provides methods for producing protein molecular weight ladders for use in protein gel electrophoresis, as well as proteins and protein molecular weight ladders produced by these methods.
REFERENCES:
patent: 4405720 (1983-09-01), Merril
patent: 4468466 (1984-08-01), Morrissey
patent: 4507233 (1985-03-01), Saito et al.
patent: 4575452 (1986-03-01), Lee et al.
patent: 4677196 (1987-06-01), Rausch et al.
patent: 4766224 (1988-08-01), Rausch
patent: 4782027 (1988-11-01), Lee et al.
patent: 4920059 (1990-04-01), Moeremans et al.
patent: 5132439 (1992-07-01), Schultz et al.
patent: 5270181 (1993-12-01), McCoy et al.
patent: 5273906 (1993-12-01), Shultz et al.
patent: 5279792 (1994-01-01), Moeremans et al.
patent: 5292646 (1994-03-01), McCoy et al.
patent: 5330902 (1994-07-01), Keck et al.
patent: 5449758 (1995-09-01), Hartley
patent: 5605691 (1997-02-01), Carroll
patent: 5616502 (1997-04-01), Haugland et al.
patent: 5705649 (1998-01-01), Shultz et al.
patent: 01-503117 (1989-10-01), None
patent: 05-507209 (1993-10-01), None
patent: 08-509504 (1996-10-01), None
patent: WO-88/07086 (1988-09-01), None
patent: WO92/01707 (1992-02-01), None
patent: WO-92/13955 (1992-08-01), None
patent: WO-95/19993 (1995-07-01), None
patent: WO-96/08570 (1996-03-01), None
patent: WO96/17942 (1996-06-01), None
patent: WO-96/30514 (1996-10-01), None
patent: WO97/28248 (1997-08-01), None
Allen, R.C., and Budowle, B., “Component Visualization,” inGel Electrophoresis of Proteins and Nucleic Acids, Walter De Gruyter, Berlin, Germany, pp. 204-272 (1994).
Ausubel, F.M., et al., “Protein Expression,” inCurrent Protocols in Molecular Biology, vol. 2, John Wiley & Sons, Inc., Hoboken, NJ, pp. 16.4.1-16.8.14 (1994).
Barger, B.O., et al., “Estimation of Molecular Weight by Polyacrylamide Gel Electrophoresis Using Heat Stable Fluorophors,”Anal. Biochem, 70:327-335, Academic Press (1976).
Betton, J-M., and Hofnung, M., “Folding of a Mutant Maltose-binding Protein ofEscherichia coliWhich Forms Inclusion Bodies,”J. Biol. Chem. 271:8046-8052, American Society for Biochemistry and Molecular Biology (1996).
BIORAD, “Electrophoresis and Blotting Standards. Protein Standards,”Life Science Research Products, Price List 5, p. 316, (1993).
Bosshard, H.F., and Datyner, A., “The Use of a New Reactive Dye for Quantitation of Prestained Proteins on Polyacrylamide Gels,”Anal. Biochem, 82:327-333, Academic Press (1977).
Buchner, J., and Rudolph, R., “Renaturation, Purification and Characterization of Recombinant FA-Fragments Produced inEscherichia coli,” Biotechnology 9:157-162, Nature Pub. Co. (1991).
Carter, P., “Site-Specific Proteolysis of Fusion Proteins,” inProtein Purification: From Molecular Mechanisms to Large-Scale Processes, American Chemical Society Symposium Series, vol. 427, Ladisch, M.R., et al., eds., American Chemical Society, Washington, DC, pp. 181-193 (1990).
Chatterjee, D.K., et al., “Cloning and overexpression of the gene encoding bacteriophage T5 DNA polymerase,”Gene 97:13-19, Elsevier Science (1991).
Chatterjee, D.K., et al., “Genetic organization of theKpn1restriction-modification system,”Nucl. Acids Res. 19:6505-6509, Oxford University Press (1991).
Coburn, G.A., and Mackie, G.A., “Overexpression, Purification, and Properties ofEscherichia coliRibonuclease II,”J. Biol. Chem. 271:1048-1053, American Society for Biochemistry and Molecular Biology (1996).
Dalmia, B.K., et al., “Domain E ofBacillus maceransCyclodextrin Glucanotransferase: An Independent Starch-Binding Domain,”Biotechnol. Bioeng. 47:575-584, Wiley Interscience (1995).
Datyner, A., and Finaimore, E.D., “A Prestaining Method for the Quantitative Assay of Proteins on Polyacrylamide Gels,”Anal. Biochem. 55:479-491, Academic Press (1973).
Dekker, N., et al., “In vitro folding ofEscherichia coliouter-membrane phospholipase A,”Eur. J. Biochem. 232:214-219, Blackwell Science (1995).
Derman, A.L , and Beckwith, J., “Escherichia coliAlkaline Phosphatase Localized to the Cytoplasm Slowly Acquires Enzymatic Activity in Cells Whose Growth Has Been Suspended: a Caution for Gene Fusion Studies,”J. Bacteriol. 177:3764-3770, American Society for Microbiology (1995).
Flynn, E., et al., “Protein Analysis with the BenchMark™ Protein Ladders,”Focus 19:33-35, Life Technologies (1997).
Georgiou, G., et al., “Localization of Inclusion Bodies inEscherichia coliOverproducing β-Lactamase or Alkaline Phosphatase,”Appl. Environ. Microbiol. 52:1157-1161, American Society for Microbiology (1986).
Griffith, I.P., “Immediate Visualization of Proteins in Dodecyl Sulfate-Polyacrylamide Gels by Prestaining with Remazol Dyes,”Anal. Biochem. 46:402-412, Academic Press (1972).
Haugland, R, “Handbook of Fluorescent Probes and Research Chemicals. Section 1.5, Dyes with Absorption Maxima Between 500 and 540 nm,” pp. 25-29, Molecular Probes Inc., available at: http://web.archive.org/web/19970607210142/www.probes.com/acrobat/ch01-5.pdf.
Haugland, R. “Handbook of Fluorescent Probes and Research Chemicals. Section 1.6, Long-Wavelength Dyes,” pp. 29-35, Molecular Probes Inc., available at: http://web.archive.org/web/19970607210142/www.probes.com/acrobat/ch01-6.pdf.
Hitomi, Y., et al., “High Efficiency Prokaryotic Expression and Purification of a Portion of the Hepatitis C Core Protein and Analysis of the Immune Response to Recombinant Protein in BALB/c Mice,”Virol Immunol. 8:109-119, Mary Ann Liebert Inc. (1995).
Hochuli, E., and Piesecki, S., “Interaction of Hexahistidine Fusion Proteins with Nitrilotriacetic Acid-Chelated Ni7+Ions,”METHODS: A Companion to Methods in Enzymology 4:68-72, Academic Press (1992).
Höög, J-O., et al., “Nucleotide sequence of the thioredoxin gene fromEscherichia coli,” Biosci. Rep. 4:917-923, The Biochemical Society (1984).
Kim, S-O., and Lee, Y.I., “High-level expression and simple purification of recombinant human insulin-like growth factor I,”J. Biotechnol 48:97-105, Elsevier Science (1996).
Kuhelj, R., et al., “The preparation of catalytically active human cathepson B from its precursor expressed inEscherichia coliin the form of inclusion bodies,”Eur. J. Biochem. 229:533-539, Blackwell Science (1995).
LaVallie, E.R., et al., “A Thioredoxin Gene Fusion Expression System That Circumvents Inclusion Body Formation in theE. coliCytoplasm,”Biotechnology 11:187-193, Nature Pub. Co. (1993).
Lunn, C.A., et al., “Amplification and Purification of Plasmid-encoded Thioredoxin fromEscherichia coliK12,”J. Biol. Chem 259:10469-10474, American Society of Biological Chemists Inc. (1984).
Magin, T.M., et al., “Analysis of cytokeratin domains by cloning and expression of intact and deleted polypeptides inEscherichia coli,” EMBO J. 6:2607-2615, IRL Press Ltd. (1987).
M
Chatterjee Deb K.
Flynn Elizabeth
Longo Mary C.
Oberfelder Robert W.
Invitrogen Corporation
Monshipouri Maryam
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