Protease-deficient cells

Details Protease-deficient cells Protease-deficient cells

2005-07-26

2005-07-26

Saidha, Tekchand (Department: 1652)

Chemistry: molecular biology and microbiology

Micro-organism, per se ; compositions thereof; proces of...

Bacteria or actinomycetales; media therefor

C435S252330, C435S226000, C435S212000, C435S069100, C435S041000, C536S023200, C530S412000

Reexamination Certificate

active

06921659

ABSTRACT:
A gram-negative bacterial cell is described that is deficient in a chromosomal gene present in a wild-type such cell which gene shares at least 80% sequence identity with the native sequence of the yfcK gene and encodes an aminopeptidase. Alternatively, a gram-negative bacterial cell is deficient in a chromosomal gene present in a wild-type such cell which gene encodes an aminopeptidase that shares at least 80% sequence identity with the native sequence of aminopeptidase b2324. Either of these types of cells, when comprising a nucleic acid encoding a heterologous polypeptide, produces an N-terminal unclipped polypeptide when it is cultured and the polypeptide recovered, with virtually no N-terminal clipped polypeptide produced as an impurity. Conversely, a method is provided for cleaving an N-terminal amino acid from a polypeptide comprising contacting the polypeptide with an aminopeptidase sharing at least 80% sequence identity with the native sequence of aminopeptidase b2324.

REFERENCES:
patent: 4865974 (1989-09-01), Ben-Bassat et al.
patent: 4946783 (1990-08-01), Beckwith et al.
patent: 5304472 (1994-04-01), Bass et al.
patent: 5508192 (1996-04-01), Georgiou et al.
patent: 5639635 (1997-06-01), Joly et al.
patent: 5789199 (1998-08-01), Joly et al.
patent: 6428997 (2002-08-01), Lee et al.
patent: 2002044601 (2002-06-01), None
patent: WO 86/01229 (1986-02-01), None
patent: WO 88/05821 (1988-08-01), None
patent: WO 00/66761 (2000-11-01), None
Alberts et al., “Normal Genes can be Easily Replaced by Mutant Ones in Bacteria and Some Lower Eucaryotes”Molecular Biology of the Cell, 3rd edition, New York:Garland Publishing, Inc. pps. 325-326 (1994).
Ausubel et al., “Chapter 16: Protein Expression”Current Protocols in Molecular Biology(Online), John Wiley & Sons, Inc. (2002).
Blattner et al., “The Complete Genome ofEscherichia coliK-12”Science(Database SwissProt on GenCore, Compugen, Ltd. GenBank, Gene Sequence) 277:1453-1474 (Sep. 1997).
Anderson et al., “Metabolic Oscillations in anE. coliFermentation.”Biotechnol. Bioeng. 75(2) :212-218 (Oct. 2001).
Bachmann., “Derivations and Genotypes of Some Mutant Derivatives ofEscherichia coliK-12.”Escherichia coli and Salmonella Typhimurium: Cellular and Molecular Biology. (Washington, DC: American Society for Microbiology.), Chapter 72, 2:1190-1219 (1987).
Bachmann., “Pedigrees of Some Mutant Strains ofEscherichia coliK-12.”Bact. Rev. 36(4) :525-557 (Dec. 1972).
Baneyx and Georgiou., “Expression of Proteolytically Sensitive Polypeptides inEscherichia coli.” Stability of Protein Pharmaceuticals., Ahern and Manning, eds., NY:Plenum Press, Chapter 3, pps. 69-108.
Blattner et al., “The Complete Genome Sequence ofEscherichia coliK-12.”Science277:1453-1462 (Sep. 1997).
Chang et al., “High-Level Secretion of Human Growth Hormone byEscherichia coli.” Gene. 55:189-196 (1987).
Chaudhury and Smith., “Escherichia colirecBC Deletion Mutants.”J. Bacteriol. 160(2) : 788-791 (Nov. 1984).
Elish et al., “Biochemical Analysis of Spontaneous fepA Mutants ofEscherichia coli.”J. Gen. Microbiol. 134:1355-1364 (1988).
Gonzales and Robert-Baudouy., “Bacterial Aminopeptidases: Properties and Functions.”FEMS Microbiology Reviews18(4) :319-344 (1996).
Joly et al., “Overexpression ofEscherichia coliOxidoreductases Increases Recombinant Insulin-Like Growth Gactor-I Accumulation.”Proc. Natl. Acad. Sci. USA95:2773-2777 (Mar. 1998).
Kleckner et al., “Genetic Engineering in Vivo Using Translocatable Drug-Resistance Elements: New Methods in Bacterial Genetics.”J. Mol. Biol. 116:125-159 (1977).
Knappik et al., “The Effect of Folding Catalysts on the In Vivo Folding Process of Different Antibody Fragments Expressed inEscherichia coli”Bio/Technology11:77-83 (Jan. 1993).
Lawther et al., “Molecular Basis of Valine Resistance inEscherichia coliK-12.”Proc. Natl. Acad. Sci. USA78:922-925 (Feb. 1981).
Mark et al., “Genetic Mapping of trxA, a Gene Affecting Thioredoxin inEscherichia coliK12.”Mol. Gen. Genet. 155:145-152 (1977).
Metcalf et al., “Use of the rep Technique for Allele Replacement to Construct NewEscherichia coliHosts for Maintenance of R6Kγ Origin Plasmids at Different Copy Numbers.”Gene. 138:1-7 (1994).
Miller, Charles G., “Protein Degradation and Proteolytic Modification.”Escherichia coli and Salmonella., Frederick C. Neidhardt, ASM Press, Chapter 62, pps. 938-954 (1996).
Park et al., “Secretory Production of Recombinant Protein by a High Cell Density Culture of a Protease Negative Mutant Escherichia coli Strain.”Biotechnol. Prog. 15:164-167 (1999).
Wolfing and Pluckthun, “Correctly Folded T-Cell Receptor Fragments in the Periplasm ofEscherichia coli”J. Mol. Biol. 242:655-669 (1994).

Affiliated with

Also associated with

Rating

Protease-deficient cells does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Protease-deficient cells, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Protease-deficient cells will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3409115