C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
195/1.22, 195/1.
C12N 15/00 (2006.01) C12N 9/52 (2006.01) C12P 21/00 (2006.01)
Patent
CA 1297051
Expression of the Cloned Lysostaphin Gene Abstract The present invention provides recombinant plasmids which in 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 pRGS and members of Bacillus species, including B. subtilis and B. sphaericus transformed by pJP1, pDF8 and pRP1 produce lysostaphin which is immunologically and elec- trophoretically 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 post-translationally processed to mature lysostaphin.
534626
Applied Microbiology Inc.
G. Ronald Bell & Associates
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