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通过使用含有大片段内部缺失的供体实现酿酒酵母交配型基因的同宗转换

Homothallic switching of Saccharomyces cerevisiae mating type genes by using a donor containing a large internal deletion.

作者信息

Weiffenbach B, Haber J E

出版信息

Mol Cell Biol. 1985 Aug;5(8):2154-8. doi: 10.1128/mcb.5.8.2154-2158.1985.

Abstract

Homothallic switching of the mating type genes of Saccharomyces cerevisiae occurs by a gene conversion event, replacing sequences at the expressed MAT locus with a DNA segment copied from one of two unexpressed loci, HML or HMR. The transposed Ya or Y alpha sequences are flanked by homologous regions that are believed to be essential for switching. We examined the transposition of a mating type gene (hmr alpha 1-delta 6) which contains a 150-base-pair deletion spanning the site where the HO endonuclease generates a double-stranded break in MAT that initiates the gene conversion event. Despite the fact that the ends of the cut MAT region no longer share homology with the donor hmr alpha 1-delta 6, switching of MATa or MAT alpha to mat alpha 1-delta 6 was efficient. However, there was a marked increase in the number of aberrant events, especially the formation of haploid-inviable fusions between MAT and the hmr alpha 1-delta 6 donor locus.

摘要

酿酒酵母交配型基因的同宗转换通过基因转换事件发生,用从两个未表达位点(HML或HMR)之一复制的DNA片段替换表达的MAT位点处的序列。转座的Ya或Yα序列两侧是被认为对转换至关重要的同源区域。我们研究了一个交配型基因(hmrα1 - δ6)的转座,该基因包含一个150个碱基对的缺失,跨越HO内切核酸酶在MAT中产生双链断裂从而启动基因转换事件的位点。尽管切割后的MAT区域末端不再与供体hmrα1 - δ6共享同源性,但MATa或MATα向matα1 - δ6的转换仍然有效。然而,异常事件的数量显著增加,特别是MAT与hmrα1 - δ6供体位点之间形成单倍体不可育融合体的情况。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e729/366935/02b95ecbb846/molcellb00104-0356-a.jpg

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