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与酵母质粒2μm环相关的复制和重组功能。

Replication and recombination functions associated with the yeast plasmid, 2 mu circle.

作者信息

Broach J R, Hicks J B

出版信息

Cell. 1980 Sep;21(2):501-8. doi: 10.1016/0092-8674(80)90487-0.

Abstract

By examining both the transformation efficiency of yeast of various plasmids containing defined regions of the 2 mu circle genome and the characteristics of the resultant transformants, we have identified several regions of the 2 mu circle genome which are involved in 2 mu circle replication or recombination. First, by identifying those DNA fragments from the molecule which promote high frequency transformation of yeast, we have localized the origin of replication to a sequence partially within the large unique region, which, as determined by subsequent deletion analysis, extends from the middle of the inverted repeat region into the contiguous unique region. Second, by examining the relative efficiency of replication in yeast of hybrid plasmids containing either the entire 2 mu circle genome or a fragment of 2 mu circle encompassing the origin of replication, we have determined that efficient use of the 2 mu circle origin requires some function or functions encoded in the molecule at a site away from the origin. Third, by examining the ability of a mutant 2 mu circle molecule to undergo intramolecular recombination in yeast, we have identified a 2 mu circle gene which codes for a product required for this process.

摘要

通过检测含有2μm环基因组特定区域的各种质粒对酵母的转化效率以及所得转化子的特征,我们确定了2μm环基因组中几个与2μm环复制或重组有关的区域。首先,通过鉴定该分子中能促进酵母高频转化的那些DNA片段,我们已将复制起点定位到部分位于大的单一区域内的一个序列,后续缺失分析确定该区域从反向重复区域的中部延伸至相邻的单一区域。其次,通过检测含有整个2μm环基因组或包含复制起点的2μm环片段的杂交质粒在酵母中的相对复制效率,我们确定有效利用2μm环起点需要该分子中远离起点的某个位点编码的一种或多种功能。第三,通过检测突变的2μm环分子在酵母中进行分子内重组的能力,我们鉴定出一个2μm环基因,该基因编码此过程所需的一种产物。

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