Lee G S, Savage E A, Ritzel R G, von Borstel R C
Department of Genetics, University of Alberta, Edmonton, Canada.
Mol Gen Genet. 1988 Nov;214(3):396-404. doi: 10.1007/BF00330472.
A forward mutation system has been developed to obtain rapidly clonable mutants at the URA3 locus in yeast by means of selection for 5-fluoroorotic acid resistance. We have used this system to determine base changes in 35 spontaneous and 34 ultraviolet radiation-induced ura3 base substitution mutants. Other mutants (frameshift, deletion, duplication, replacement) were detected as well. Evidence is reported which suggests cyclobutane dimers are the principal mutagenic lesions induced by UV radiation in stationary phase cells of the yeast Saccharomyces cerevisiae. Since most of the induced lesions are at 5'-TT-3' sites, the results suggest that the "A-rule", preferential insertion of adenine residues opposite poorly pairing sites in DNA, does not apply for yeast cells irradiated in stationary phase, whereas the spontaneous mutation data indicate that the A-rule applies for cells in logarithmic phase. Most of the spontaneous mutations are transversions. UV-induced transitions and transversions occur at approximately equal frequencies.
已开发出一种正向突变系统,通过选择对5-氟乳清酸具有抗性,来快速获得酵母URA3基因座处的可克隆突变体。我们已使用该系统来确定35个自发的和34个紫外线辐射诱导的ura3碱基替换突变体中的碱基变化。还检测到了其他类型的突变体(移码、缺失、重复、替换)。有证据表明,环丁烷二聚体是酿酒酵母静止期细胞中紫外线辐射诱导的主要诱变损伤。由于大多数诱导损伤位于5'-TT-3'位点,结果表明,“A规则”(即在DNA中与配对不佳的位点相对优先插入腺嘌呤残基)不适用于静止期照射的酵母细胞,而自发突变数据表明A规则适用于对数期的细胞。大多数自发突变是颠换。紫外线诱导的转换和颠换发生频率大致相等。