Molecular cloning of the Shv-1 beta-lactamase gene and construction of an Shv-1 hybridization probe.
We have cloned the Shv-1 beta-lactamase gene from the R1010 plasmid into pACYC184. By subcloning and transposon mutagenesis we have localized the gene to a 1.6 kb BscI-SalI fragment of R1010, which is present in the recombinant plasmid pUB8. A 900 bp PstI fragment of pUB8 was shown to be a specific hybridization probe by testing against plasmids which encode 17 different beta-lactamase enzymes. A comparison was made of the sensitivity of the Shv-1 probe labelled with either [35S]dCTP or with photobiotin.
Bisessar
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School of Biological Sciences, University of East Anglia, Norfolk, UK.
James
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D003001:Cloning, Molecular; D015342:DNA Probes; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005819:Genetic Markers; D009154:Mutation; D009693:Nucleic Acid Hybridization; D001618:beta-Lactamases / Q000235:genetics*
D015342:DNA Probes; D004251:DNA Transposable Elements; D005819:Genetic Markers; D001618:beta-Lactamases