Kataoka T, Powers S, McGill C, Fasano O, Strathern J, Broach J, Wigler M
Cell. 1984 Jun;37(2):437-45. doi: 10.1016/0092-8674(84)90374-x.
We present a genetic analysis of RAS1 and RAS2 of S. cerevisiae, two genes that are highly homologous to mammalian ras genes. By constructing in vitro ras genes disrupted by selectable genes and introducing these by gene replacement into the respective ras loci, we have determined that neither RAS1 nor RAS2 are by themselves essential genes. However, ras1 - ras2 - spores of doubly heterozygous diploids are incapable of resuming vegetative growth. We have determined that RAS1 is located on chromosome XV, 7 cM from ade2 and 63 cM from his3; and RAS2 is located on chromosome XIV, 2 cM from met4 . We have also constructed by site-directed mutagenesis a missense mutant, RAS2val19 , which encodes valine in place of glycine at the nineteenth amino acid position, the same sort of missense mutation that is found in some transforming alleles of mammalian ras genes. Diploid yeast cells that contain this mutation are incapable of sporulating efficiently, even when they contain wild-type alleles.
我们对酿酒酵母的RAS1和RAS2进行了遗传分析,这两个基因与哺乳动物的ras基因高度同源。通过构建被选择基因破坏的体外ras基因,并通过基因替换将其引入各自的ras基因座,我们确定RAS1和RAS2本身都不是必需基因。然而,双杂合二倍体的ras1 - ras2 - 孢子无法恢复营养生长。我们确定RAS1位于第十五号染色体上,距ade2为7厘摩,距his3为63厘摩;RAS2位于第十四号染色体上,距met4为2厘摩。我们还通过定点诱变构建了一个错义突变体RAS2val19,它在第十九位氨基酸位置编码缬氨酸而非甘氨酸,这是在一些哺乳动物ras基因的转化等位基因中发现的同类型错义突变。含有这种突变的二倍体酵母细胞即使含有野生型等位基因也无法有效形成孢子。