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对酵母整合转化进行高效紫外线刺激需要质粒两条链都受损。

Efficient UV stimulation of yeast integrative transformation requires damage on both plasmid strands.

作者信息

Ninković M, Alacević M, Fabre F, Zgaga Z

机构信息

Faculty of Food and Biotechnology, University of Zagreb, Croatia.

出版信息

Mol Gen Genet. 1994 May 10;243(3):308-14. doi: 10.1007/BF00301066.

Abstract

The nature of UV-induced pre-recombinational structures was studied using transformation of Saccharomyces cerevisiae cells with non-replicative plasmids. Transformation by double-stranded plasmids irradiated with UV was stimulated up to 50-fold, and both plasmid integration and conversion of the mutated chromosomal selective gene were found to be equally increased. The stimulation observed with such 'totally' irradiated plasmids was not found with plasmids bearing lesions in only one strand. This effect is attributed to the formation by excision repair of recombinogenic structures consisting of a pyrimidine dimer opposite a gap. When single-stranded integrative plasmids were irradiated, their transforming potential was decreased but the proportion of transformants that arose by gene conversion, rather than by plasmid integration, was increased from 8% to 49% as a function of the UV dose. Possible reasons why single-strand UV lesions favour gene conversion are discussed.

摘要

利用非复制性质粒对酿酒酵母细胞进行转化,研究了紫外线诱导的重组前结构的性质。用紫外线照射的双链质粒转化可被刺激高达50倍,并且发现质粒整合和突变的染色体选择基因的转化均同样增加。在仅一条链上带有损伤的质粒中未发现用这种“完全”照射的质粒所观察到的刺激作用。这种效应归因于通过切除修复形成由与缺口相对的嘧啶二聚体组成的重组结构。当单链整合质粒受到照射时,其转化潜力降低,但通过基因转化而非质粒整合产生的转化体比例随紫外线剂量的增加从8%提高到49%。讨论了单链紫外线损伤有利于基因转化的可能原因。

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