Construction by homologous recombination and phenotypic characterization of a DNA polymerase domain polA mutant of Mycobacterium smegmatis.
Gene replacement was achieved by homologous recombination in Mycobacterium smegmatis (Ms) using a cloned segment of the polA gene (encoding the DNA polymerase I) disrupted within the region encoding the C-terminal DNA polymerase domain by a kanamycin-resistance marker. The Ms polA755:aph mutant thus generated displayed a phenotype of hypersensitivity to DNA damage induced by UV irradiation and by hydrogen peroxide challenge.
Gordhan
B G
BG
Molecular Biology Unit, South African Institute for Medical Research, Johannesburg, South Africa.
Andersen
S J
SJ
De Meyer
A R
AR
Mizrahi
V
V
D003001:Cloning, Molecular; D004249:DNA Damage; D004256:DNA Polymerase I / Q000235:genetics* / Q000378:metabolism; D018390:Gene Targeting; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D009161:Mycobacterium / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000528:radiation effects; D010641:Phenotype; D011995:Recombination, Genetic; D015183:Restriction Mapping
D006861:Hydrogen Peroxide; D004256:DNA Polymerase I