Vector-mediated DNA double-strand break repair analysis in normal, and radiation-sensitive, Chinese hamster V79 cells.
DNA double-strand break repair was assessed in 2 new radiation-sensitive V79 hamster cell lines (irs1 and irs2) by their ability to rejoin restriction endonuclease cuts in a transferred selectable SV40--E. coli gpt recombinant gene. The studied gene was carried in the vector pPMH16 which also contained a second selectable HSVtk-neo recombinant gene which acted as a control for DNA transformation. The parental V79 cells showed correct rejoining of KpnI and EcoRV double-strand breaks in approximately 18% and 36% of transformants respectively (correcting for the expression of undamaged gpt in neo+ transformants). irs1 shows a significantly reduced (approximately 3-fold) ability to rejoin correctly such double-strand scissions. However, irs2 rejoined such lesions as correctly as the V79 cells. The data are discussed in the context of the assay and the possible repair deficiencies of these radiosensitive mutant cells.
Debenham
P G
PG
Division of Cell and Molecular Biology, MRC Radiobiology Unit, Didcot, Oxon, Great Britain.
Jones
N J
NJ
Webb
M B
MB
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004260:DNA Repair; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004274:DNA, Recombinant; D005822:Genetic Vectors; D011839:Radiation, Ionizing; D014170:Transformation, Genetic
D004274:DNA, Recombinant; D004262:DNA Restriction Enzymes