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沙门氏菌中染色体区域相互之间的不均等可及性:利用噬菌体λ整合酶介导的长程重排探究染色体结构

Unequal access of chromosomal regions to each other in Salmonella: probing chromosome structure with phage lambda integrase-mediated long-range rearrangements.

作者信息

Garcia-Russell Nathalie, Harmon Timothy G, Le Tien Q, Amaladas Nelusha H, Mathewson Richard D, Segall Anca M

机构信息

Department of Biology and Center for Microbial Sciences, San Diego State University, 5500 Campanile Dr. LS416, San Diego, CA 92182-4614, USA.

出版信息

Mol Microbiol. 2004 Apr;52(2):329-44. doi: 10.1111/j.1365-2958.2004.03976.x.

Abstract

We have investigated the fluidity of the Salmonella chromosome architecture using the phage lambda site-specific recombination system as a probe. We determined how chromosome position affects the extent of integrase-mediated recombination between pairs of inversely oriented att sites at various loci. We also investigated the accessibility of each chromosomal att site to an extrachromosomal partner carried on a low-copy plasmid. Recombination events were assayed by semi-quantitative polymerase chain reaction of the attP product. The extent of recombination between the chromosome and the plasmid was generally higher than intrachromosomal recombination except for two loci, araA::attL and galT::attL, which gave no detectable recombination with any other locus. Based on 20 intervals, we found that chromosomal locations are not equally accessible to each other. Although multiple factors probably affect accessibility, the most important is the specific combination of the end-points used. Neither the size of the intervals nor the accessibility of individual end-points to extrachromosomal sequences is as important. These results suggest that the chromosome is not completely fluid but rather organized in some way, with barriers that limit the movement of DNA within the cell. The nature of the barriers involved in chromosomal organization remains to be determined.

摘要

我们使用噬菌体λ位点特异性重组系统作为探针,研究了沙门氏菌染色体结构的流动性。我们确定了染色体位置如何影响整合酶介导的位于不同位点的反向att位点对之间的重组程度。我们还研究了每个染色体att位点与低拷贝质粒携带的染色体外伴侣的可及性。通过对attP产物进行半定量聚合酶链反应来检测重组事件。除了两个位点araA::attL和galT::attL与任何其他位点均未检测到重组外,染色体与质粒之间的重组程度通常高于染色体内重组。基于20个区间,我们发现染色体位置之间的可及性并不相同。尽管可能有多种因素影响可及性,但最重要的是所使用端点的特定组合。区间大小和单个端点对染色体外序列的可及性都没有那么重要。这些结果表明,染色体并非完全流动,而是以某种方式组织起来的,存在限制细胞内DNA移动的屏障。参与染色体组织的屏障的性质仍有待确定。

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