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[转座子δTn9'中由IS1元件介导的共整合质粒的结构与稳定性]

[Structure and stability of cointegrating plasmids mediated by the IS1 elements in transposon delta Tn9'].

作者信息

Danilevich V N, Alkhimova R A, Rebentish B A, Sukhodolets V V

出版信息

Genetika. 1990 Jan;26(1):18-29.

PMID:2160901
Abstract

In order to elucidate the structural features of the transposon Tn9', representative of the Tn9 family, which define the ability of the transposon to produce unstable cointegrates, we have obtained a derivative of this transposon carrying a deletion in its central region. The deletion in the obtained transposon delta Tn9' covers a DNA segment of about 50 bp in length, occupying the most distal position in relation to the cat gene, at its junction with the right copy of the IS1. The structure and stability of the IS1/delta Tn9'-mediated cointegrates between the plasmids pDK57.1 (pBR322::delta Tn9') and pRP3.1, a deletion derivative of RP1, have been studied. The three types of cointegrates were found. Those of the type I are predominantly formed, due to the left copy of the IS1 which in delta Tn9' occupies proximal position to the promoter of the cat gene. These cointegrates contain three copies of IS1 and are of high stability. The cointegrates of the type II contain two entire copies of delta Tn9' (i.e. four copies of IS1) as well as the structures of the type II, representing the cointegrate equivalent of inverse transposition and also containing four copies of IS1. Cointegrates of the type II and III dissociate efficiently in the rec+ cells but, in contrast to the cointegrates mediated by the original transposon Tn9', are unable to dissociate efficiently in the recA- cells. It was concluded that a DNA segment in the central region of Tn9' may be essential for the expression of the IS1-specific resolvase encoded by the right copy of IS1.

摘要

为了阐明转座子Tn9'(Tn9家族的代表)的结构特征,这些特征决定了转座子产生不稳定共合体的能力,我们获得了该转座子的一个衍生物,其中心区域存在缺失。所获得的转座子δTn9'中的缺失覆盖了一段长度约为50 bp的DNA片段,该片段在与IS1右拷贝的连接处,相对于cat基因处于最远端位置。我们研究了质粒pDK57.1(pBR322::δTn9')和RP1的缺失衍生物pRP3.1之间由IS1/δTn9'介导的共合体的结构和稳定性。发现了三种类型的共合体。I型共合体主要是由于IS1的左拷贝在δTn9'中位于cat基因启动子的近端位置而形成的。这些共合体包含三个IS1拷贝,具有高稳定性。II型共合体包含两个完整的δTn9'拷贝(即四个IS1拷贝)以及II型结构,代表反向转座的共合体等效物,也包含四个IS1拷贝。II型和III型共合体在rec+细胞中能有效解离,但与原始转座子Tn9'介导的共合体不同,在recA-细胞中不能有效解离。得出的结论是,Tn9'中心区域的一个DNA片段可能对于由IS1右拷贝编码的IS1特异性解离酶的表达至关重要。

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