Expression of the cloned lysostaphin, gene.

摘要

Expression of the Cloned Lysostaphin GeneAbstractThe present invention provides recombinantplasmids which in transformant microbial hosts expresslysostaphin, a bacteriocin that kills most knownstaphylococcal species. The invention also provideslysostaphin, substantially free from non-lysostaphincontaminants. Recombinant plasmids, pRG5, pJP1, pDF8and pRP1, were derived by inserting a 1.5 kilobasesegment of DNA coding for lysostaphin into the cloningvectors, pUC8, pBC16, pBD64 and pSPV1, respectively.E. coli strain JM105 transformed by pRGS and membersof Bacillus species, including B. subtilis andB. sphaericus transformed by pJP1, pDF8 and pRP1produce lysostaphin which is immunologically and elec-trophoretically indistinguishable from that producedby S. simulans, the natural source. Furthermore,B sphaericus strain 00/pJP1 transformants produce fivetimes the amount of lysostaphin as S. simulans. Theinvention also provides the 1.5 kbp DNA fragmentcoding for lysostaphin. The sequence of the DNAencodes preprolysostaphin, a monomeric 389 amino acidprotein, which is post-translationally processed tomature lysostaphin.