Open AccessUse of a novel allele of the Escherichia coli aacC4 aminoglycoside resistance gene as a genetic marker in mycobacteria
Author(s) -
Consaul Sandra A.,
Pavelka Martin S.
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2004.tb09547.x
Subject(s) - kanamycin , biology , aminoglycoside , escherichia coli , genetics , plasmid , allele , gene , microbiology and biotechnology , mutant , mutagenesis , antibiotics
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.