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首页> 外文期刊>Journal of bacteriology >Characterization of conjugative plasmid EDP208.
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Characterization of conjugative plasmid EDP208.

机译：接合质粒EDP208的表征。

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摘要

EDP208 is a conjugative plasmid belonging to incompatibility group IncF0 lac, A restriction endonuclease map of this plasmid was constructed using five restriction enzymes: BamHI, HindIII, PvuI, SstI, and XhoI. On the basis of these mapping studies, the plasmid was found to be 90 kilobases in length. Clones were constructed from four large HindIII fragments of plasmid EDP208. One fragment, HindIII-20.5, was found to contain the lac genes and the origin of vegetative replication (oriV). Another fragment, HindIII-27.5, was found to contain all of the genes necessary for sex pilus formation, but it was nontransmissible. However, when used to complement a plasmid carrying an adjacent fragment, HindIII-23, the transfer of the latter occurred, suggesting that HindIII-23 contains the origin of transfer (oriT). The further localization of genes concerned with pilus biosynthesis was achieved by transposon mutagenesis. Six EDP208::Tn1 and thirty-seven EDP208::Tn5 mutants were isolated on the basis of their resistance to f1, a filamentous phage which adheres to intact pilus tips. The positions of the inserted transposons were determined on the restriction map and a 16.5-kilobase region was found to be required for pilus synthesis.
机译：EDP​​208是属于不相容性组IncF0 lac的结合质粒，使用5种限制酶BamHI，HindIII，PvuI，SstI和XhoI构建了该质粒的限制性核酸内切酶图。在这些作图研究的基础上，发现质粒的长度为90kb。从质粒EDP208的四个大HindIII片段构建克隆。发现一个片段，HindIII-20.5，包含lac基因和营养性复制（oriV）的起源。发现另一个片段，HindIII-27.5，包含性菌毛形成所需的所有基因，但它是不可传播的。但是，当用于与携带相邻片段HindIII-23的质粒互补时，发生了后者的转移，这表明HindIII-23包含转移起点（oriT）。转座子诱变实现了菌毛生物合成相关基因的进一步定位。根据它们对f1的抗性，分离了6个EDP208 :: Tn1和37个EDP208 :: Tn5突变体，f1是一种附着于完整菌毛末端的丝状噬菌体。在限制性酶切图上确定插入的转座子的位置，发现菌毛合成需要16.5碱基的区域。

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