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Journal of Antimicrobial Chemotherapy (1994) 34, 715-725
© 1994 The British Society for Antimicrobial Chemotherapy

research-article

Nucleotide sequence and genetic analysis of the type Ib trimethoprim-resistant, Tn4132-encoded dihydrofolate reductase

Hilary-Kay Young, M. J. Qumsieh and M. Louise Mclntosh

Department of Biological Sciences, University of Dundee Dundee DD1 4HN, UK

Received 1 November 1993; accepted 15 February 1994

Transposon Tn4132, encoding the type Ib trimethoprim-resistant, dihydrofolate reductase, was transposed from the clinical plasmid pUK163 to a small recombinant plasmid pZMR12. Restriction endonuclease and partial sequence analysis of Tn4132 revealed a close relationship to Tn7. The nucleotide sequence of the dhfrlb trimethoprim-resistance gene was determined and the gene and its product were found to share significant homology with the dhfrla, V, VI and VII plasmid-encoded dihydrofolate reductases. Extensive sequence homology (88%) was observed with the type V dihydrofolate reductase at both the nucleotide and amino acid level. Oligonucleotide probes, distinguishing between the dhfrlb and dhfrV genes, were designed. The discriminatory capabilities of these probes in future epidemiological studies will permit a more accurate determination of the dissemination of these two closely related trimethoprim-resistance genes than has previously been possible.


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