Alenin V V, Getsova M L
Genetika. 1997 Jun;33(6):858-61.
A simple technique of selection for spontaneous red mutants of yeast Saccharomyces cerevisiae is proposed. It is based on incubation of yeast cells carrying a deletion in the GCN4 gene on a solid selective medium containing 3-amino-1, 2, 4-triazole (3AT). After incubation for three weeks at 30 degrees C, the 3 AT-resistant mutants ade2 and ade1 were easily visualized due to the pink or red colony color; they were then selected and analyzed by genetic and biochemical methods. The frequency of these colored mutants was approximately 10-9. The method proposed is shown to be suitable for selection and analysis of both spontaneous mutants ade2 and ade1 and those induced by various mutagens. The possible molecular mechanism of 3AT-resistance of red mutants is discussed. This mechanism involves GCN4-independent activation of expression of genes of purine and histidine metabolism. This activation results from purine starvation caused by arrest of purine biosynthesis the ade2 and ade2 mutations.
本文提出了一种筛选酿酒酵母自发红色突变体的简单技术。该技术基于将携带GCN4基因缺失的酵母细胞在含有3-氨基-1,2,4-三唑(3AT)的固体选择培养基上培养。在30℃培养三周后,由于菌落呈粉红色或红色,ade2和ade1这两种抗3AT的突变体很容易被观察到;然后对它们进行选择,并通过遗传和生化方法进行分析。这些有色突变体的频率约为10-9。结果表明,所提出的方法适用于自发突变体ade2和ade1以及各种诱变剂诱导的突变体的筛选和分析。文中讨论了红色突变体对3AT抗性的可能分子机制。该机制涉及嘌呤和组氨酸代谢基因的GCN4非依赖性表达激活。这种激活是由ade2和ade2突变导致嘌呤生物合成停滞引起的嘌呤饥饿所致。