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利用缺口填补法直接克隆酵母基因组及酿酒酵母第六条染色体的完整物理图谱

The direct cloning of the yeast genome using the gap-filling method and the complete physical mapping of Saccharomyces cerevisiae chromosome VI.

作者信息

Iwasaki T, Shirahige K, Yoshikawa H, Ogasawara N

机构信息

Department of Genetics, Osaka University Medical School, Japan.

出版信息

Gene. 1991 Dec 20;109(1):81-7. doi: 10.1016/0378-1119(91)90591-x.

Abstract

The ordered clone library of chromosome VI of Saccharomyces cerevisiae has been constructed by Olson et al. [Proc. Natl. Acad. Sci. USA 83 (1986) 7826-7830, and personal communication]. It is composed of four contiguous stretches from the chromosome, each of 40-70 kb. There remained three gaps of unknown length between these four contigs. We applied the 'gap-repair' method to clone these three gap regions directly from the yeast chromosome. All three gap regions, ranging from 7 to 22 kb, were successfully cloned without any structural changes. Together with these gap regions, a precise physical map of EcoRI and HindIII sites was constructed over the 230-kb fragment which covers most of chromosome VI except for two telomeres.

摘要

酿酒酵母第六条染色体的有序克隆文库已由奥尔森等人构建[《美国国家科学院院刊》83 (1986) 7826 - 7830,以及个人交流]。它由来自该染色体的四个连续片段组成,每个片段长度为40 - 70 kb。这四个重叠群之间仍存在三个长度未知的间隙。我们应用“缺口修复”方法直接从酵母染色体克隆这三个间隙区域。所有三个间隙区域,长度在7至22 kb之间,均成功克隆,且无任何结构变化。连同这些间隙区域,在覆盖除两个端粒外大部分第六条染色体的230 kb片段上构建了EcoRI和HindIII位点的精确物理图谱。

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