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酿酒酵母1号染色体DNA的分子克隆:ADE1基因的分离

Molecular cloning of chromosome I DNA from Saccharomyces cerevisiae: isolation of the ADE1 gene.

作者信息

Crowley J C, Kaback D B

出版信息

J Bacteriol. 1984 Jul;159(1):413-7. doi: 10.1128/jb.159.1.413-417.1984.

Abstract

The ADE1 gene of Saccharomyces cerevisiae was isolated by complementation in S. cerevisiae from a yeast genomic DNA library carried on plasmid YEp13. Electron microscopy of R-loop-containing DNA indicated the location of the ADE1 gene on the plasmid insert. Gene disruption and gene replacement were used to demonstrate that the ade1-complementing sequence was the actual ADE1 gene that maps on chromosome I. ade1 strains which normally form red colonies form white ones when transformed with the cloned ADE1 gene. This property should be very useful, since it enables detection of plasmids carrying this gene under nonselective conditions.

摘要

通过在酿酒酵母中对携带于质粒YEp13上的酵母基因组DNA文库进行互补作用,分离出了酿酒酵母的ADE1基因。对含R环的DNA进行电子显微镜观察,确定了ADE1基因在质粒插入片段上的位置。利用基因破坏和基因置换技术证明,ade1互补序列就是位于第I号染色体上的实际ADE1基因。通常形成红色菌落的ade1菌株,在用克隆的ADE1基因转化后会形成白色菌落。这一特性应该非常有用,因为它能够在非选择性条件下检测携带该基因的质粒。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0e75/215651/5ed1543901a1/jbacter00230-0422-a.jpg

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