Vector to produce biologically important peptides

Details Vector to produce biologically important peptides Vector to produce biologically important peptides

1991-06-14

1995-03-21

Wax, Robert A.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435 694, 4353201, 4352528, 536 241, 935 47, 935 48, C12N 1517, C12N 1516, C12N 1570, C12N 1563

Patent

active

053994906

ABSTRACT:
In this patent application we have described the construction of a novel secretion vector based on E. coli enterotoxin coding sequence. We have shown categorically that pre and pro regions of toxin gene are absolutely necessary for extra cellular secretion of the stable toxin. We have also shown with specific examples that when the nucleotide coding sequence of a heterologous peptide is fused in frame to the end of the pro region in the st gene, the resultant vector in an E. coli host secretes extracellularly correctly processed heterologous peptide. This application also includes construction of suitable vectors where this gene fusion can be achieved. Generally methods to create such fusions involving a) recombinant DNA technology and b) the use of site directed in vitro mutagenesis, have also been described. A general method of purification of heterologous peptides is also described in this application. This novel vector system can be used for hyperproduction and extracellular secretion of peptides of biological importance.

REFERENCES:
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Guzman-Verduzco et al., Infection & Immunity 57(2):645-648 (1989).
Okamoto, K. et al. J. of Bacteriology 172(9):5260-5265 (1990).
Dwarakanath, P. et al., Gene 81:219-226 (1989).
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Stieglitz, H. et al., Plasmid 20:42-53 (1988).
Suggs, S. et al. "Use of Synthetic Oligonucleotides as Hybridization Probes." PNAS 78(11)6613-6617 (1981).
Marston, F. A. O. "The Purification of Eukaryotic Polypeptides Synthesized in E. coli." Biochem J. 240:1-12 (1986).

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