Expression of the cloned lysostaphin gene

Details Expression of the cloned lysostaphin gene Expression of the cloned lysostaphin gene

1987-04-10

1990-06-05

Martinell, James

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

435 691, 435220, 4351723, 435243, 4352523, 43525231, 43525233, 43525235, 435255, 435256, 435320, 536 27, 935 11, 935 29, 935 69, 935 72, 935 73, 935 74, 935 75, C12N 900, C12N 2102, C12N 2100, C12N 1500, C12N 100, C12N 120, C12N 116, C12N 118, C07H 2104

Patent

active

049313908

ABSTRACT:
The present invention provides recombinant plasmids which is transformant microbial hosts express lysostaphin, a bacteriocin that kills most known staphylococcal species. The invention also provides lysostaphin, substantially free from non-lysostaphin contaminants. Recombinant plasmids, pRG5, pJP1, pDF8 and pRP1, were derived by inserting a 1.5 kilobase segment of DNA coding for lysostaphin into the cloning vectors, pUC8, pBC16, pBD64 and pSPV1, respectively. E. coli strain JM105 transformed by pRG5 and members of Bacillus species, including B. subtilis and B. sphaericus transformed by pJP1, pDF8 and pRP1 produce lysostaphin which is immunologically and electrophoretically indistinguishable from that produced by S. simulans, the natural source. Furthermore, B. sphaericus strain 00/pJP1 transformants produce five times the amount of lysostaphin as S. simulans. The invention also provides the 1.5 kbp DNA fragment coding for lysostaphin. The sequence of the DNA encodes preprolysostaphin, a monomeric 389 amino acid protein, which is posttranslationally processed to mature lysostaphin.

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Paul A. Recsei, "Cloning, Sequence, and Expression of the Lysostaphin Gene from Staphylococcus Simulans"; Proc. Natl. Acad. Sci. U.S.A., vol. 84, pp. 1127-1131, Mar. 1987 Biochemistry.
L. S. Heath et al., "Expression in Escherichia coli of the Gene Encoding Lysostaphin Endopeptidase," (Abstr.) 87th Annual Meeting of the American Society for Microbiology, 1987.

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