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pSC101质粒par区域的结构与功能分析

Structural and functional analysis of the par region of the pSC 10 1 plasmid.

作者信息

Tucker W T, Miller C A, Cohen S N

出版信息

Cell. 1984 Aug;38(1):191-201. doi: 10.1016/0092-8674(84)90540-3.

Abstract

Three distinct segments (the partition-related, or PR segments) within the 370 bp par region of pSC101 have been shown by deletion analysis to be involved in partitioning of the plasmid to daughter cells. The two lateral segments are direct repeats, each of which potentially can pair with an inverted repeat located between them to form a hairpin-loop structure. Deletion of either lateral segment, together with the middle segment, results in plasmid instability (the Par- phenotype). Deletion of one PR segment yields a stable plasmid that nevertheless shows reduced ability to compete with a coexisting wild-type derivative of the same replicon (the Cmp- phenotype). Deletion of all three segments results in a rate of plasmid loss far in excess of that predicted from the observed copy number of the plasmid. Analysis of the segregation properties of these mutants and of temperature-sensitive and high copy number derivatives of the pSC101 replicon suggests a model in which the par function allows the nonreplicating plasmids of the intracellular pool to be counted as individual molecules, and to be distributed evenly to daughter cells. In the absence of par, the multicopy pool of plasmids behaves as a single segregation unit.

摘要

通过缺失分析表明,pSC101的370 bp par区域内有三个不同的片段(与分配相关的,或PR片段)参与质粒向子细胞的分配。两个侧翼片段是直接重复序列,每个都可能与位于它们之间的反向重复序列配对形成发夹环结构。删除任何一个侧翼片段以及中间片段都会导致质粒不稳定(Par-表型)。删除一个PR片段会产生一个稳定的质粒,但其与同一复制子的共存野生型衍生物竞争的能力会降低(Cmp-表型)。删除所有三个片段会导致质粒丢失率远远超过根据观察到的质粒拷贝数预测的水平。对这些突变体以及pSC101复制子的温度敏感型和高拷贝数衍生物的分离特性分析表明,存在一种模型,其中par功能允许细胞内质粒池中未复制的质粒被视为单个分子,并均匀地分配到子细胞中。在没有par的情况下,多拷贝质粒池表现为单个分离单位。

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