Use of a novel allele of the Escherichia coli aacC4 aminoglycoside resistance gene as a genetic marker in mycobacteria.
The aacC4 gene from Escherichia coli can be expressed in mycobacteria and confers resistance to apramycin. However, the major limitation of the aacC4 gene as a genetic tool is that the gene also confers resistance to kanamycin and gentamicin, two antibiotics commonly used for selection in mycobacterial genetics, thus reducing the utility of the aacC4 gene in the mycobacterial field. To overcome this problem we constructed, by chemical mutagenesis, a mutant allele of the E. coli aacC4 gene that still confers resistance to apramycin but has a reduced ability to confer resistance to kanamycin and gentamicin. We then constructed a variety of E. coli-mycobacteria shuttle plasmids containing this mutant allele.
Consaul
Sandra A
SA
Department of Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, 601 Elmwood Avenue, P.O. Box 672, Rochester, NY 14642, USA.
Pavelka
Martin S
MS
D000617:Aminoglycosides / Q000494:pharmacology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics*; D005819:Genetic Markers; D005839:Gentamicins / Q000494:pharmacology; D007612:Kanamycin / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D016297:Mutagenesis, Site-Directed; D009161:Mycobacterium / Q000235:genetics*; D009328:Nebramycin / Q000031:analogs & derivatives* / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D015183:Restriction Mapping
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D005819:Genetic Markers; D005839:Gentamicins; C487816:aacC4 protein, E coli; D009328:Nebramycin; C011666:apramycin; D007612:Kanamycin