Nucleic acid hybridization analysis of an integrated plasmid in Staphylococcus aureus

Abstract

A series of studies were performed on a Staphylococcus aureus strain thought to contain a pencillinase plasmid integrated into the host chromosome. Reassociation kinetics analysis of whole-cell deoxyribonucleic acid (DNA) in the presence of pure radioactive plasmid DNA revealed that plasmid-specific sequences were present at about 1 copy per chromosome equivalent as compared to 3.6 copies for the same plasmid in its autonomous state. Consistent with this observation was the finding that penicillinase activity was lower for the former strain than for the latter. It was shown further that the plasmid-specific sequences cosedimented on neutral sucrose gradients with fragments of whole-cell DNA many times larger than the plasmid. These two findings were taken as strongly confirmatory of the integrated state. Analysis of whole-cell ribonucleic acid for the presence of plasmid-specific messengers revealed that these were present in approximately the amounts expected on the basis of the DNA study.