Cloning and expression in Enterobacteriaceae of the extended-spectrum beta-lactamase gene from a Pseudomonas aeruginosa plasmid.
An extended-spectrum beta-lactamase, the gene for which is located on plasmid pMS350 in Pseudomonas aeruginosa strains, hydrolyzes carbapenems and other extended-spectrum beta-lactam antibiotics. We cloned the pMS350 beta-lactamase gene in an Escherichia coli K-12 strain using the vector plasmid pHSG398, and subcloned it into pMS360, a plasmid with a wide host-range. This resulted in the formation of the recombinant plasmid, pMS363, containing a 4.1-kb DNA insert that includes the extended-spectrum beta-lactamase gene. Plasmid pMS363 was introduced into the P. aeruginosa PAO strain or into six species of Enterobacteriaceae, and the specific activities of the beta-lactamase and MICs of various beta-lactam antibiotics were estimated. The cloned gene was capable of expression in these strains and caused resistance to carbapenem, penem and other beta-lactam antibiotics, with the exception of aztreonam.
Iyobe
S
S
Laboratory of Drug Resistance in Bacteria, Gunma University School of Medicine, Maebashi, Japan.
Tsunoda
M
M
Mitsuhashi
S
S
D003001:Cloning, Molecular; D004755:Enterobacteriaceae / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D010957:Plasmids; D011550:Pseudomonas aeruginosa / Q000235:genetics*; D001618:beta-Lactamases / Q000235:genetics*
D001618:beta-Lactamases