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来自质粒R1卡那霉素抗性区域的一段140个碱基对的DNA片段充当复制起点并促进位点特异性重组。

A 140 base-pair DNA segment from the kanamycin resistance region of plasmid R1 acts as an origin of replication and promotes site-specific recombination.

作者信息

Clerget M

出版信息

J Mol Biol. 1984 Sep 5;178(1):35-46. doi: 10.1016/0022-2836(84)90229-8.

Abstract

A 140 base-pair DNA segment situated just upstream of the kanamycin resistance gene of transposon Tn2350, a transposon carried by the plasmid R1, was found to act as an origin of replication and allow autonomous replication of a plasmid composed only of the segment and of the tetracycline resistance gene of pBR322. This segment also promotes site-specific recombination: when cloned in pBR322 it promotes multimer formation in a recA- strain. If two copies are cloned on the same plasmid they promote either deletion or inversion of the intervening region, depending on their orientation relative to each other. DNA gyrase seems to be involved in this process since the inversion rate, in a plasmid carrying sequences in opposite orientations, varies in different nalidixic acid-resistant strains (gyr A mutants) independently isolated.

摘要

位于转座子Tn2350(一种由质粒R1携带的转座子)卡那霉素抗性基因上游的一段140个碱基对的DNA片段,被发现可作为复制起点,并允许仅由该片段和pBR322的四环素抗性基因组成的质粒进行自主复制。该片段还能促进位点特异性重组:当克隆到pBR322中时,它能促进recA-菌株中多聚体的形成。如果两个拷贝克隆在同一质粒上,它们会根据彼此的方向促进中间区域的缺失或倒位。DNA促旋酶似乎参与了这一过程,因为在携带相反方向序列的质粒中,倒位率在独立分离的不同萘啶酸抗性菌株(gyr A突变体)中有所不同。

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