RT Journal Article SR Electronic(1) A1 Phonimdaeng, Prasart A1 O’Reilly, Mary A1 O’Toole, Paul W. A1 Foster, Timothy J.YR 1988 T1 Molecular Cloning and Expression of the Coagulase Gene of Staphylococcus aureus 8325-4 JF Microbiology, VO 134 IS 1 SP 75 OP 83 DO https://doi.org/10.1099/00221287-134-1-75 PB Microbiology Society, SN 1465-2080, AB The gene coding for coagulase (coa) was cloned from Staphylococcus aureus 8325-4 in a λ replacement vector in Escherichia coli. Coagulase (plasma-clotting) activity was measured in λ coa lysates and an immunoreactive protein of 60 kDa was detected by Western immunoblotting with anti-coagulase serum. This protein comigrated with the major immunoreactive protein in supernatants of S. aureus 8325-4. The coa gene was subcloned in pUC vectors. One recombinant expressed a 60 kDa immunoreactive protein and plasma-clotting activity. A putative β-galactosidase-coagulase fusion protein and truncated peptides were expressed by variants formed by subcloning. These results are consistent with previously published biochemical data that the prothrombin-binding domain of coagulase is located in the N-terminus of the protein. The cloned coa gene was transferred into S. aureus on a shuttle plasmid. Expression of coagulase was higher in a strain with a mutation in the agr locus, which controls the level of several exoproteins in S. aureus, suggesting that agr normally regulates coagulase expression negatively., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-134-1-75