Transfer of a plasmid-specified beta-lactamase gene from Haemophilus influenzae

Abstract

A number of ampicillin-resistant strains of Haemophilus influenzae could donate a gene specifying the type IIIa (TEM) beta-lactamase to Haemophilus parainfluenzae, Escherichia coli, and Pseudomonas aeruginosa. Donor strains rapidly lost their ability to transfer ampicillin resistance on storage or subculture. Such strains also apparently contained a single species of covalently closed circular deoxyribonucleic acid of contour length 1.2 mum, equivalent to about 2.5 x 10(6) daltons. No species of plasmid deoxyribonucleic acid large enough to encode sex factor activity was detected. Despite this, transfer occurred to several bacterial genera in the presence of deoxyribonuclease, suggesting that transmissibility was by conjugation. The beta-lactamase gene was generally unstable after transfer and was lost in the absence of selection. Where stable transcipients were found, this was evidently by insertion of the beta-lactamase gene into the host chromosome. In P. aeruginosa insertion was always accompanied by induction of auxotrophy for adenine, suggesting insertion at a specific site. It is believed that insertion also occurred at one site on the chromosome of Escherichia coli. Crypticity measurements for beta-lactamase activity showed that there was little or no penetration barrier to beta-lactam drugs in Haemophilus. This may explain the long delay in the acquisition of ampicillin resistance by this organism.