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酿酒酵母RAD50基因的克隆与定位

Cloning and mapping of the RAD50 gene of Saccharomyces cerevisiae.

作者信息

Kupiec M, Simchen G

出版信息

Mol Gen Genet. 1984;193(3):525-31. doi: 10.1007/BF00382094.

Abstract

The RAD50 gene was cloned as a 4.8 kb fragment in the 2 mu derived plasmid pFL1. The gene resides in a 3.9 kb segment that was subcloned into the plasmid YRp7. The cloned gene complements the deficiency caused by the rad50-1 mutation with respect to gamma-rays, MMS resistance and UV-induced mitotic recombination. Restoration of the Rad+ phenotype occurs when the cloned gene is on a freely replicating multiple-copy plasmid or in the integrated form. Mapping of the cloned gene following integration of the 2 mu plasmid, and of the subclone in plasmid YRp7, showed it to be located on the left arm of chromosome XIV. Tetrad analysis of various crosses involving two different strains carrying rad50-1 showed the mutation to map next to pet2 on chromosome XIV, and not on the right arm of chromosome IV, as previously published.

摘要

RAD50基因作为一个4.8 kb的片段克隆于源自2μm的质粒pFL1中。该基因位于一个3.9 kb的片段上,此片段被亚克隆到质粒YRp7中。克隆的基因在γ射线、对甲磺酸甲酯(MMS)的抗性以及紫外线诱导的有丝分裂重组方面,弥补了rad50 - 1突变所导致的缺陷。当克隆的基因存在于自由复制的多拷贝质粒上或以整合形式存在时,Rad +表型得以恢复。对2μm质粒整合后以及质粒YRp7中的亚克隆进行克隆基因定位,结果表明它位于第十四号染色体的左臂上。对涉及携带rad50 - 1的两种不同菌株的各种杂交组合进行四分体分析,结果显示该突变位于第十四号染色体上pet2旁边,而非如先前发表的位于第四号染色体的右臂上。

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