Fukui Y, Kaziro Y
EMBO J. 1985 Mar;4(3):687-91. doi: 10.1002/j.1460-2075.1985.tb03684.x.
We have cloned a ras gene homologue from fission yeast Schizosaccharomyces pombe and determined its nucleotide sequence. A putative coding sequence for 219 amino acids was found. The sequence contained one set of splicing signals: GTAAGT for a donor sequence, ACTAA for a unique sequence found in introns of yeast genes and TAG for an acceptor sequence, indicating the existence of an intron. The amino-terminal one third of the predicted S. pombe ras protein was nearly perfectly homologous and the next one third moderately homologous to those of mammalian ras proteins. The carboxy-terminal one third showed no homology but terminated with a short conserved sequence Cys-X-X-Z (X being a hydrophobic amino acid) as in other ras proteins. The result of Southern analysis of S. pombe DNA under nonstringent hybridization conditions using our clone as a probe indicated that no other closely related gene may be present in the S. pombe genome. The transcript of this gene could be detected by Northern analysis.
我们从裂殖酵母粟酒裂殖酵母中克隆了一个ras基因同源物,并测定了其核苷酸序列。发现了一个推测的编码219个氨基酸的序列。该序列包含一组剪接信号:供体序列为GTAAGT,酵母基因内含子中独特序列为ACTAA,受体序列为TAG,表明存在一个内含子。预测的粟酒裂殖酵母ras蛋白的氨基末端三分之一与哺乳动物ras蛋白的氨基末端三分之一几乎完全同源,接下来的三分之一与哺乳动物ras蛋白的氨基末端三分之一中度同源。羧基末端三分之一没有同源性,但像其他ras蛋白一样以短保守序列Cys-X-X-Z(X为疏水氨基酸)结尾。在非严格杂交条件下,用我们的克隆作为探针进行粟酒裂殖酵母DNA的Southern分析结果表明,粟酒裂殖酵母基因组中可能不存在其他密切相关的基因。通过Northern分析可以检测到该基因的转录本。