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酿酒酵母RAD18基因的分离及rad18缺失突变体的构建。

Isolation of the RAD18 gene of Saccharomyces cerevisiae and construction of rad18 deletion mutants.

作者信息

Fabre F, Magana-Schwencke N, Chanet R

机构信息

Institut Curie-Biologie, Centre Universitaire, Orsay, France.

出版信息

Mol Gen Genet. 1989 Feb;215(3):425-30. doi: 10.1007/BF00427039.

Abstract

The RAD18 gene of Saccharomyces cerevisiae is involved in mutagenic DNA repair. We describe its isolation from a yeast library introduced into the centromeric YCp50 vector, a low copy number plasmid. The insert was subcloned into YCp50 and into the multicopy YRp7 plasmid. RAD18 is not toxic when present in multiple copies but the UV survival response indicates an heterogeneity in the cell population, a fraction of it being more sensitive. A DNA segment, close to RAD18, is toxic on the multicopy plasmid and may correspond to the tRNA sup61 known to be tightly linked to RAD18. Chromosomal deletions of RAD18 were constructed. The gene is not essential and the deleted strains have the properties of single site mutants. Thus, RAD18 appears to be essentially involved in DNA repair metabolism.

摘要

酿酒酵母的RAD18基因参与诱变DNA修复。我们描述了从引入着丝粒YCp50载体(一种低拷贝数质粒)的酵母文库中分离该基因的过程。该插入片段被亚克隆到YCp50和多拷贝YRp7质粒中。RAD18基因多拷贝存在时无毒,但紫外线存活反应表明细胞群体存在异质性,其中一部分细胞更敏感。靠近RAD18的一个DNA片段在多拷贝质粒上具有毒性,可能对应于已知与RAD18紧密连锁的tRNA sup61。构建了RAD18基因的染色体缺失突变体。该基因并非必需基因,缺失该基因的菌株具有单点突变体的特性。因此,RAD18似乎主要参与DNA修复代谢。

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