SELECTION OF SPONTANEOUS MUTANTS BY INOSITOL STARVATION IN YEAST
SELECTION OF SPONTANEOUS MUTANTS BY INOSITOL STARVATION IN YEAST
复制标题
DOI：
10.1007/bf00269415
复制
发表时间：
1975-01-01
期刊：
影响因子：
--
通讯作者：
CULBERTSON, MR
中科院分区：
文献类型：
--
作者：
HENRY, SA;DONAHUE, TF;CULBERTSON, MR
报道了一种新的常规分离酵母（Saccharomycescerevisiae）自发突变的方法。该技术是基于肌醇营养缺陷型细胞在缺乏肌醇时死亡的观察。然而，如果宏分子合成被抑制，大多数细胞存活。因此，对肌醇需要突变体的适当操作可导致具有影响大分子合成的突变的细胞的选择性存活。由于回复到肌醇原养型可能是干扰有效选择的主要来源，因此已经构建了单倍体双突变株，其回复到肌醇原养型的频率估计比单个自发突变事件的预期频率低几个数量级。使用这种菌株，富集超过10000倍，已获得各类营养缺陷型突变体。还获得了自发的温度敏感突变体。
A new method for the routine isolation of mutations of spontaneous origin in the yeast,Saccharomyces cerevisiae, is reported. The technique is based on the observation that inositol auxotrophs die when deprived of inositol. However, if marromolecular synthesis is inhibited, most of the cells survive. Appropriate manipulation of inositol requiring mutants can threfore result in the selective survival of cells possesing mutations which affect macromolecular synthesis. Since reversion to inositol prototrophy can be a major source of interference in efficient selection, a haploid double mutant strain has been constructed which reverts to inositol prototrophy with a frequency estimated to be several orders of magnitude lower than the expected frequency of single, spontaneous mutational events. Using this strain, enrichment in excess of 10000 fold has been obtained for various classes of auxotrophic mutants. Spontaneous temperature sensitive mutants have also been obtained.