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A cryptic 65-kilobase-pair transposonlike element isolated from Bacteroides uniformis has homology with Bacteroides conjugal tetracycline resistance elements.

机译:从均匀的拟杆菌中分离出的一种秘密的65碱基对转座子样元件与拟杆菌结合的四环素抗性元件具有同源性。

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摘要

A 65-kilobase-pair element, XBU4422, which has some transposonlike characteristics but carries no known antibiotic resistance genes, has been isolated from Bacteroides uniformis 0061. XBU4422 was trapped on Bacteroides-Escherichia coli shuttle vectors during experiments in which one of the conjugal Bacteroides tetracycline resistance (Tcr) elements was being used to mobilize the shuttle vectors to Bacteroides recipients. Results of Southern hybridization experiments showed that XBU4422 is normally integrated in the B. uniformis 0061 chromosome and is found only in some strains. Insertion of XBU4422 in the shuttle vectors was site specific and orientation specific. Nonmobilizable vectors that had acquired XBU4422 became transmissible and could be transferred to Bacteroides or E. coli recipients. In B. uniformis transconjugants, the XBU4422 insertion in the vectors was usually intact, but XBU4422 was always lost in matings with E. coli, Bacteroides thetaiotaomicron, or B. ovatus. The loss of XBU4422 did not visibly alter the vector; in the case of E. coli, the loss of the insertion appeared to be RecA dependent. Although XBU4422 carried no antibiotic resistances, it shared regions of homology with six conjugal Bacteroides Tcr elements; this homology was strongest with the ends of XBU4422. Using a strain of B. thetaiotaomicron that contains no XBU4422-hybridizing sequences, we showed that the ends of XBU4422 were probably reacting with the ends of the Tcr elements. These results provide the first direct evidence that the Tcr elements, like XBU4422, are integrated in the chromosome and that insertion of the least some Tcr elements, such as TcrEmr DOT, is relatively site specific.
机译:已从uniformactides homois 0061中分离出具有65个碱基对的元件XBU4422,该元件具有一些类似转座子的特征,但不携带已知的抗生素抗性基因。XBU4422在实验过程中被捕获在细菌-大肠杆菌穿梭载体上,在该实验中,一种共生细菌使用四环素抗性(Tcr)元件将穿梭载体动员到拟杆菌的受体上。 Southern杂交实验的结果表明,XBU4422通常整合在统一芽孢杆菌0061染色体中,仅在某些菌株中发现。 XBU4422在穿梭载体中的插入是位点特异性和方向特异性的。已获得XBU4422的不可动载体变得可以传播,可以转移到拟杆菌或大肠杆菌受体中。在统一芽孢杆菌转导结合体中,载体中的XBU4422插入通常是完整的,但XBU4422总是在与大肠杆菌,拟杆菌(Bacteroides thetaiotaomicron)或卵形芽孢杆菌的交配中丢失。 XBU4422的丢失没有明显改变载体。在大肠杆菌的情况下,插入的丢失似乎是RecA依赖性的。尽管XBU4422不具有抗生素抗性,但它与六个结合的拟杆菌属Tcr元件具有同源性。 XBU4422的末端具有最强的同源性。使用不包含XBU4422-杂交序列的thetaiotaomicron菌株,我们显示XBU4422的末端可能与Tcr元件的末端反应。这些结果提供了第一个直接证据,表明Tcr元件(如XBU4422)已整合到染色体中,并且至少一些Tcr元件(如TcrEmr DOT)的插入相对位点特异性。

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