Rapid detection of a specific trimethoprim resistance gene using a biotinylated DNA probe

A DNA probe specific for the dihydrofolate reductase (DHFR) type I gene was labelled with biotin by the process of nick-translation and used to screen 83 independently-derived trimethoprim R plasmids from Enterobacteriaceae. Hybridization was detected using streptavidine and a biotin-conjugated alka...

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Bibliographic Details
Published in:Journal of antimicrobial chemotherapy 1987-09, Vol.20 (3), p.335-341
Main Authors: Carter, G. I., Towner, K. J., Slack, R. C. B.
Format: Article
Language:English
Subjects:
Alkaline Phosphatase - metabolism
Bacteriology
Biological and medical sciences
Biotin - metabolism
DNA - analysis
Fundamental and applied biological sciences. Psychology
Genetics
Microbiology
Nucleic Acid Hybridization
Plasmids
Tetrahydrofolate Dehydrogenase - genetics
Trimethoprim Resistance - genetics
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Summary:A DNA probe specific for the dihydrofolate reductase (DHFR) type I gene was labelled with biotin by the process of nick-translation and used to screen 83 independently-derived trimethoprim R plasmids from Enterobacteriaceae. Hybridization was detected using streptavidine and a biotin-conjugated alkaline phosphatase to generate an insoluble coloured precipitate following the addition of an appropriate dye. Sixty-eight plasmids (81.9%) hybridized with the probe for DHFR type I. The method could be adapted for use with any antibiotic resistance gene for which a suitable DNA probe is available and has none of the drawbacks associated with the use of radioactively-labelled DNA in hybridization techniques.
ISSN:0305-7453
1460-2091
DOI:10.1093/jac/20.3.335