Nagpal M L, Higgins D R, Prakash S
Mol Gen Genet. 1985;199(1):59-63. doi: 10.1007/BF00327510.
The RAD1 and RAD3 genes of Saccharomyces cerevisiae are required for excision repair of UV damaged DNA. In addition, the RAD3 gene is essential since rad3 deletions are recessive lethals. We have examined the induction of the RAD1 and RAD3 genes by DNA damage and during the cell division cycle. We have made fusions of the RAD1 and RAD3 genes with the Escherichia coli lacZ gene encoding beta-galactosidase. Beta-galactosidase activity was measured in a Rad+ yeast strain containing the RAD1-lacZ or the RAD3-lacZ fusion, either in a multicopy replicating plasmid or as a single copy integrant resulting from transformation with an integrating plasmid which transforms yeast by homologous recombination in the yeast genome. No induction of beta-galactosidase activity occurred after ultraviolet light (UV) or 4-nitroquinoline-1-oxide (NQO) treatment. Haploid cells of mating type a were synchronized by treatment with alpha factor and beta-galactosidase activity was determined during different cell cycle stages. No change in beta-galactosidase activity was observed in the strain containing the RAD1-lacZ or the RAD3-lacZ fusion integrated in the yeast genome.
酿酒酵母的RAD1和RAD3基因是紫外线损伤DNA切除修复所必需的。此外,RAD3基因是必需的,因为rad3缺失是隐性致死的。我们研究了DNA损伤以及细胞分裂周期中RAD1和RAD3基因的诱导情况。我们构建了RAD1和RAD3基因与编码β-半乳糖苷酶的大肠杆菌lacZ基因的融合体。在含有RAD1-lacZ或RAD3-lacZ融合体的Rad+酵母菌株中测量β-半乳糖苷酶活性,该融合体存在于多拷贝复制质粒中,或者是由整合质粒转化酵母基因组后通过同源重组产生的单拷贝整合体。紫外线(UV)或4-硝基喹啉-1-氧化物(NQO)处理后,未诱导β-半乳糖苷酶活性。通过用α因子处理使a交配型单倍体细胞同步化,并在不同细胞周期阶段测定β-半乳糖苷酶活性。在酵母基因组中整合了RAD1-lacZ或RAD3-lacZ融合体的菌株中,未观察到β-半乳糖苷酶活性的变化。