CLONING AND CHARACTERIZATION OF THE RAD9 DNA-REPAIR GENE FROM SCHIZOSACCHAROMYCES POMBE
CLONING AND CHARACTERIZATION OF THE RAD9 DNA-REPAIR GENE FROM SCHIZOSACCHAROMYCES POMBE
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DOI：
10.1093/nar/19.13.3525
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发表时间：
1991-07-11
影响因子：
14.9
通讯作者：
WATTS, FZ
中科院分区：
文献类型：
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作者：
MURRAY, JM;CARR, AM;WATTS, FZ
rad9。来自分裂酵母的DNA修复突变体裂糖酵母对紫外线和电离辐射都很敏感。通过对突变细胞系γ射线敏感性的互补，克隆出了rad9基因。一个4.3kb的HindIII片段被发现对这两种类型的辐射具有抵抗力。互补区域进一步定位到2.6kb的HindIII-EcoRV片段，通过DNA序列分析，发现该片段包含能够编码427个氨基酸的蛋白质的序列，如果假设三个内含子去除停止密码子。内含子通过cDNA克隆序列分析和cDNA衍生的PCR产物得到证实。转录产物为1.6kb的低丰度mRNA。推测的rad9蛋白与任何已发表的序列没有同源性。截断的蛋白能够补充rad9的辐射敏感性。192突变体。该基因的缺失不是致命的，空等位基因具有与rad9.192突变体相似的表型。
The rad9. 192 DNA repair mutant from the fission yeast, Schizosaccharomyces pombe, is sensititive to both UV and ionising radiation. The rad9 gene has been cloned by complementation of the gamma-ray sensitivity of the mutant cell line. A 4.3kb HindIII fragment was found to confer resistance to both types of radiation. The region of complementation was further localised to a 2.6kb HindIII-EcoRV fragment, which, by DNA sequence analysis, was found to contain sequences capable of coding for a 427 amino acid protein, if three introns were postulated to remove stop codons. The introns were confirmed by sequence analysis of cDNA clones and PCR products derived from cDNA. The product of transcription is a 1.6kb mRNA of low abundance. The putative rad9 protein shows no homology to any published sequence. A truncated protein is capable of complementing the radiation sensitivity of the rad9. 192 mutant. Deletion of the gene is not lethal and the null allele has a similar phenotype to the rad9.192 mutant.