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首页> 外文期刊>Journal of bacteriology >Tn7 transposition as a probe of cis interactions between widely separated (190 kilobases apart) DNA sites in the Escherichia coli chromosome.
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Tn7 transposition as a probe of cis interactions between widely separated (190 kilobases apart) DNA sites in the Escherichia coli chromosome.

机译：Tn7转座是大肠杆菌染色体中广泛分离的DNA位点（相距190 KB）之间顺式相互作用的探针。

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摘要

We have used the bacterial transposon Tn7 to examine communication between widely separated DNA sites in the Escherichia coli chromosome. Using Tn7 target immunity, a regulatory feature of transposition which influences target selection, we have evaluated (i) how the presence of Tn7 sequences at one DNA site affects Tn7 insertion into another site in the same DNA molecule and (ii) the nucleotide distances over which the two sites are able to communicate. We demonstrate that Tn7 sequences at one chromosomal site act at a distance to inhibit insertion of Tn7 elsewhere in that DNA as far away as 190 kb, reflecting effective long-range cis interactions. We have found that while target immunity is effective over a substantial region of the chromosome, insertion of Tn7 into a more distant site 1.9 Mb away in the same DNA is not inhibited; this observation provides evidence that target immunity relies on DNA spacing. We also find that within the region of the chromosome affected by target immunity, the magnitude of the immune effect is greater at close DNA sites than DNA sites farther away, suggesting that target immunity is distance dependent. We also extend the characterization of the Tn7 end-sequences involved in transposition and target immunity and describe how Tn7 target immunity can be used as a tool for probing bacterial chromosome structure.
机译：我们已经使用细菌转座子Tn7来检查大肠杆菌染色体中广泛分离的DNA位点之间的通讯。使用Tn7靶标免疫力，它是影响靶标选择的转座调控特征，我们评估了（i）一个DNA位点上Tn7序列的存在如何影响Tn7插入同一DNA分子中另一个位点，以及（ii）核苷酸距离这两个站点能够通信。我们证明一个染色体位点上的Tn7序列在远处起作用以抑制Tn7在该DNA的其他地方插入高达190 kb，反映了有效的长距离顺式相互作用。我们发现，虽然靶标免疫在整个染色体的大部分区域内都是有效的，但在同一个DNA中将Tn7插入更远的1.9 Mb的位点并没有受到抑制;该观察提供了靶免疫力依赖于DNA间隔的证据。我们还发现，在受靶标免疫力影响的染色体区域内，靠近DNA部位的免疫效果的幅度大于远处的DNA部位，表明靶标免疫力是距离依赖性的。我们还扩展了转座和靶标免疫中涉及的Tn7末端序列的特征，并描述了如何将Tn7靶标免疫用作探测细菌染色体结构的工具。

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