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首页> 外文期刊>Molecular and Cellular Biology >RAD3 gene of Saccharomyces cerevisiae: nucleotide sequence of wild-type and mutant alleles, transcript mapping, and aspects of gene regulation.
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RAD3 gene of Saccharomyces cerevisiae: nucleotide sequence of wild-type and mutant alleles, transcript mapping, and aspects of gene regulation.

机译：酿酒酵母的RAD3基因：野生型和突变型等位基因的核苷酸序列，转录本作图以及基因调控方面。

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摘要

We determined the complete nucleotide sequence of the RAD3 gene of Saccharomyces cerevisiae. The coding region of the gene contained 2,334 base pairs that could encode a protein with a calculated molecular weight of 89,796. Analysis of RAD3 mRNA by Northern blots and by S1 nuclease mapping indicated that the transcript was approximately 2.5 kilobases and did not contain intervening sequences. Fusions between the RAD3 gene and the lac'Z gene of Escherichia coli were constructed and used to demonstrate that the RAD3 gene was not inducible by DNA damage caused by UV radiation or 4-nitroquinoline-1-oxide. Two UV-sensitive chromosomal mutant alleles of RAD3, rad3-1 and rad3-2, were rescued by gap repair of a centromeric plasmid, and their sequences were determined. The rad3-1 mutation changed a glutamic acid to lysine, and the rad3-2 mutation changed a glycine to arginine. Previous studies have shown that disruption of the RAD3 gene results in loss of an essential function and is associated with inviability of haploid cells. In the present experiments, plasmids carrying the rad3-1 and rad3-2 mutations were introduced into haploid cells containing a disrupted RAD3 gene. These plasmids expressed the essential function of RAD3 but not its DNA repair function. A 74-base-pair deletion at the 3' end of the RAD3 coding region or a fusion of this deletion to the E. coli lac'Z gene did not affect either function of RAD3.
机译：我们确定了酿酒酵母RAD3基因的完整核苷酸序列。该基因的编码区包含2334个碱基对，可以编码蛋白质，计算分子量为89796。通过Northern印迹和S1核酸酶作图分析RAD3 mRNA，结果表明该转录本约为2.5kb，不包含插入序列。构建了RAD3基因和大肠杆菌的lac'Z基因之间的融合体，并用于证明RAD3基因不能被紫外线或4-硝基喹啉-1-氧化物引起的DNA损伤诱导。 RAD3的两个紫外线敏感的染色体突变等位基因rad3-1和rad3-2，通过对着丝粒质粒的缺口修复而获得，并确定了它们的序列。 rad3-1突变将谷氨酸变为赖氨酸，而rad3-2突变将甘氨酸变为精氨酸。先前的研究表明，RAD3基因的破坏导致基本功能的丧失，并与单倍体细胞的成活率有关。在本实验中，将携带rad3-1和rad3-2突变的质粒引入含有破坏的RAD3基因的单倍体细胞中。这些质粒表达了RAD3的基本功能，但没有表达其DNA修复功能。 RAD3编码区3'端的74个碱基对的缺失或该缺失与大肠杆菌lac'Z基因的融合均不影响RAD3的功能。

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