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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Cointegrate-resolution of toluene-catabolic transposon Tn4651: Determination of crossover site and the segment required for full resolution activity
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Cointegrate-resolution of toluene-catabolic transposon Tn4651: Determination of crossover site and the segment required for full resolution activity

机译：甲苯分解代谢转座子Tn4651的协整分辨率：确定交叉位点和全分辨率活性所需的片段

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摘要

Tn. 3-family transposon Tn. 4651 from Pseudomonas putida mt-2 plasmid pWW0 carries two divergently transcribed genes, tnpS and tnpT, for cointegrate-resolution. While tnpS encodes a tyrosine recombinase, tnpT encodes a protein that shows no homology to any other characterized protein. The Tn. 4651 resolution site was previously mapped within the 203-bp fragment that covered the tnpS and tnpT promoter region. To better understand the molecular mechanisms underlying the Tn. 4651 cointegrate-resolution, we determined the extent of the functional resolution site (designated the rst site) of Tn. 4651 and the location of the crossover site for the cointegrate-resolution. Deletion analysis of the rst region localized the fully functional rst site to a 136-bp segment. The analysis of the site-specific recombination between Tn. 4651 rst and a rst variant from the Tn. 4651-related transposon, Tn. 4661, indicated that the crossover occurs in the 33-bp inverted repeat region, which separates the 136-bp functional rst site into the tnpS- and tnpT-proximal segments. Electrophoretic mobility shift assays demonstrated specific binding of TnpT to the 20-bp inverted repeat region in the tnpT-proximal segment. The requirement for accessory sequences on both sides of the crossover site and the involvement of the unique DNA-binding protein TnpT suggest that the Tn. 4651-specified resolution system uses a different mechanism than other known resolution systems. Furthermore, comparative sequence analysis for Tn. 4651-related transposons revealed the occurrence of DNA exchange at the rst site among different transposons, suggesting an additional role of the TnpS-TnpT-. rst system in the evolution of Tn. 4651-related transposons.
机译：Tn。 3族转座子Tn。来自恶臭假单胞菌mt-2质粒4651的4651携带两个不同转录的基因tnpS和tnpT，以实现整体整合。虽然tnpS编码酪氨酸重组酶，但tnpT编码的蛋白质与任何其他表征的蛋白质均无同源性。 Tn。先前将4651分辨率位点定位在覆盖tnpS和tnpT启动子区域的203bp片段内。为了更好地了解Tn的分子机制。 4651共积分分辨率，我们确定了Tn的功能分辨率位点（指定为第一个位点）的范围。 4651和交叉点的位置以实现共积分分辨率。第一个区域的缺失分析将功能齐全的第一个位点定位在一个136 bp的片段上。 Tn之间的位点特异性重组的分析。 4651 rst和Tn的rst变体。 4651相关转座子，田纳西州。 4661表明，交换发生在33bp的反向重复区域中，该区域将136bp的功能性第一个位点分为tnpS和tnpT近端片段。电泳迁移率变动分析证明了TnpT与tnpT近端片段中20 bp反向重复区域的特异性结合。交叉位点两侧都需要辅助序列，而独特的DNA结合蛋白TnpT的参与提示了Tn。 4651指定的分辨率系统使用与其他已知分辨率系统不同的机制。此外，Tn的比较序列分析。 4651相关的转座子揭示了不同转座子之间在第一个位点发生了DNA交换，这暗示了TnpS-TnpT-的额外作用。 Tn演化中的第一个系统。 4651相关的转座子。

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