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[转座子Tn9'中IS1元件转座过程中形成的共整合质粒和简单插入结构的特征]

[Characteristic features of the structure of co-integrating plasmids and simple insertions formed during transposition of the IS1 element in transposon Tn9'].

作者信息

Danilevich V N, Sukhodolets V V

出版信息

Mol Biol (Mosk). 1989 Sep-Oct;23(5):1279-88.

PMID:2558290
Abstract

Earlier we have studied unstable dissociating IS1/Tn9'-mediated cointegrates between the plasmids pDK57 (pBR322::Tn9') and pRP3.1, a deletion derivative of RP1, and two types of such cointegrates containing three and four copies of IS1 were revealed. In the present paper we studied the structure of stable IS1/Tn9'-mediates cointegrates and simple insertions formed by interaction between the plasmids pDK57 and pRP3.1 in the E. coli recA- cells. It was shown, that the stable cointegrates were formed by insertion of pDK57 in different loci of pRP3.1 and these cointegrates contain three copies of IS1, i.e. one copy of IS1 and a copy of Tn9' at the junction of the two replicons. The cointegrates are formed predominantly due to the activity of the left copy of Tn9', which occupies a proximal position in regard to the promoter of the cat gene. It was found that the integration of pDK57 into the kan gene region of pRP3.1 leading to the formation of the KmS cointegrates occurs only in one of the two possible orientations. Meanwhile the insertions of the transposon Tn9' into the kan region of pRP3.1 leading to simple insertions occurs in the orientation opposite to the orientation of the transposon in the KmS cointegrates. It is proposed that simple insertions are not the products of direct transposition of Tn9', but they are formed from unstable cointegrates under the action of IS1-specific resolvase.

摘要

此前我们研究了不稳定的解离型IS1/Tn9'介导的质粒pDK57(pBR322::Tn9')和RP1的缺失衍生物pRP3.1之间的共整合体,并且揭示了两种类型的此类共整合体,它们分别含有三个和四个IS1拷贝。在本文中,我们研究了稳定的IS1/Tn9'介导的共整合体的结构以及大肠杆菌recA-细胞中质粒pDK57和pRP3.1相互作用形成的简单插入片段。结果表明,稳定的共整合体是由pDK57插入pRP3.1的不同位点形成的,这些共整合体含有三个IS1拷贝,即在两个复制子的连接处有一个IS1拷贝和一个Tn9'拷贝。共整合体主要是由于Tn9'的左拷贝的活性形成的,该拷贝相对于cat基因的启动子占据近端位置。发现pDK57整合到pRP3.1的kan基因区域导致KmS共整合体的形成仅在两种可能的方向之一中发生。同时,转座子Tn9'插入pRP3.1的kan区域导致简单插入的情况发生在与KmS共整合体中转座子方向相反的方向。有人提出,简单插入不是Tn9'直接转座的产物,而是在IS1特异性解离酶的作用下由不稳定的共整合体形成的。

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