Construction by homologous recombination and phenotypic characterization of a DNA polymerase domain polA mutant of Mycobacterium smegmatis

B G Gordhan; S J Andersen; A R De Meyer; V Mizrahi

doi:10.1016/0378-1119(96)00350-2

Construction by homologous recombination and phenotypic characterization of a DNA polymerase domain polA mutant of Mycobacterium smegmatis

Gene. 1996 Oct 31;178(1-2):125-30. doi: 10.1016/0378-1119(96)00350-2.

Authors

B G Gordhan¹, S J Andersen, A R De Meyer, V Mizrahi

Affiliation

¹ Molecular Biology Unit, South African Institute for Medical Research, Johannesburg, South Africa.

PMID: 8921903
DOI: 10.1016/0378-1119(96)00350-2

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cloning, Molecular
DNA Damage
DNA Polymerase I / genetics*
DNA Polymerase I / metabolism
Gene Targeting
Hydrogen Peroxide / pharmacology
Molecular Sequence Data
Mutation
Mycobacterium / drug effects
Mycobacterium / enzymology*
Mycobacterium / genetics
Mycobacterium / radiation effects
Phenotype
Recombination, Genetic
Restriction Mapping

Substances

Hydrogen Peroxide
DNA Polymerase I

Associated data

GENBANK/U26405