Abstract: Recombinant vectors and a method for producing a predetermined protein in a trademark microbial host are provided. The recombinant vectors are autonomously replicable expression vectors for use in B. subtilis and E. coli and express proteins therein. The vectors contain DNA sequences which code for preprolysostaphin protein secretion sequences which are necessary and sufficient for efficient secretion of the predetermined protein by transformed hosts. A gene for a predetermined protein is inserted into the vectors such that the order of genes in the vector is 5'-vector DNA-preprolysostaphin protein secretion sequence-predetermined protein sequence-vector DNA-3'. Optionally and preferably, the vectors also contain additional DNA sequences coding for staphylococcal nuclease which is a secretable detectable protein tag.

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.