Singh A, Sherman F
Genetics. 1975 Sep;81(1):75-97. doi: 10.1093/genetics/81.1.75.
One hundred and thirty-three spontaneous and induced mutants of the met15 locus in Saccharomyces cerevisiae were characterized with respect to temperature sensitivity, osmotic remediability, interallelic complementation, and suppressibility by amber and ochre suppressors. Forty mutants are osmotic remedial; 17 of these, and no others, are also temperature-sensitive. Seven of 133 mutations are suppressible by an amber suppressor and 11 are suppressible by an ochre suppressor. Seventy percent of the mutants exhibited interallelic complementation, suggesting that the functional gene product of the met15 gene is a multimeric protein. Relative map positions of 30 met15 were estimated from the frequencies of X-ray-induced mitotic reversion of various heteroallelic diploids. All complementing nonsense mutations are located near one end of the gene in contrast to other nonsense mutations which span most of the gene, thus relating the direction of translation of the mRNA with respect to the fine-structure map. Recombination studies indicated that two of 30 mutants contained deletions of the entire met15 locus. -- It was established that a variety of mutational types, including missense, nonsense, and deletions, are recovered with this unique system in which both forward and reverse mutations can be selected on the basis of methyl mercury resistance and methionine requirement of the met15 mutants.
对酿酒酵母中met15基因座的133个自发和诱导突变体进行了温度敏感性、渗透压修复性、等位基因间互补以及琥珀突变和赭石突变抑制性等方面的特征分析。40个突变体具有渗透压修复性;其中17个(且只有这17个)同时对温度敏感。133个突变中有7个可被琥珀突变抑制,11个可被赭石突变抑制。70%的突变体表现出等位基因间互补,这表明met15基因的功能性基因产物是一种多聚体蛋白。根据各种异源等位基因二倍体经X射线诱导的有丝分裂回复频率,估算了30个met15突变的相对图谱位置。与其他跨越基因大部分区域的无义突变不同,所有互补的无义突变都位于基因的一端,从而确定了mRNA的翻译方向与精细结构图谱的关系。重组研究表明,30个突变体中有2个包含整个met15基因座的缺失。——已经确定,在这个独特的系统中可以获得多种突变类型,包括错义突变、无义突变和缺失突变,在该系统中,可以根据met15突变体对甲基汞的抗性和对甲硫氨酸的需求来选择正向和反向突变。