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脆弱拟杆菌BV-17中5-硝基咪唑抗性质粒pIP417的接合转移:转移区域的特性及核苷酸序列分析

Conjugal transfer of the 5-nitroimidazole resistance plasmid pIP417 from Bacteroides vulgatus BV-17: characterization and nucleotide sequence analysis of the mobilization region.

作者信息

Trinh S, Haggoud A, Reysset G

机构信息

Unité des Anaérobies, Institut Pasteur, Paris, France.

出版信息

J Bacteriol. 1996 Dec;178(23):6671-6. doi: 10.1128/jb.178.23.6671-6676.1996.

Abstract

Three small 5-nitroimidazole (5-Ni) resistance plasmids (pIP417, pIP419, and pIP421) from Bacteroides clinical isolates are transferable by a conjugative process during homologous or heterologous matings. The mobilization properties of pIP417 originated from strain BV-17 of Bacteroides vulgatus were studied. The plasmid was successfully introduced by in vitro conjugation into different strains of Bacteroides and Prevotella species and could be transferred back from these various strains to a plasmid-free 5-Ni-sensitive Bacteroides fragilis strain, indicating that in vivo spread of the resistance gene may occur. The transfer of plasmid pIP417 harbored by the Tc(r) strain BF-2 of B. fragilis was stimulated by low concentrations of tetracycline or chlorotetracycline. This suggests a possible role for coresident conjugative transposons in the dissemination of 5-Ni resistance among gram-negative anaerobes. The nucleotide sequence of the 2.1-kb DNA mobilization region was determined. It contains a putative origin of transfer (oriT) in an A+T-rich-region, including three inverted repeats, and two integration host factor binding sites. The two identified mobilization genes (mobA and mobB) are organized in one operon and were both required for efficient transfer. Southern blotting indicated that the mobilization region of plasmid pIP417 is closely related to that of both the erythromycin resistance plasmid pBFTM1O and the 5-Ni resistance plasmid pIP419 but not to that of the 5-Ni resistance plasmid pIP421.

摘要

从拟杆菌临床分离株中获得的三个小型5-硝基咪唑(5-Ni)抗性质粒(pIP417、pIP419和pIP421)可在同源或异源交配过程中通过接合作用进行转移。对源自脆弱拟杆菌BV-17菌株的pIP417的转移特性进行了研究。该质粒通过体外接合成功导入不同的拟杆菌属和普雷沃菌属菌株,并可从这些不同菌株转移回无质粒的5-Ni敏感脆弱拟杆菌菌株,这表明抗性基因可能在体内传播。脆弱拟杆菌的Tc(r)菌株BF-2携带的质粒pIP417的转移受到低浓度四环素或金霉素的刺激。这表明共存的接合转座子在革兰氏阴性厌氧菌中5-Ni抗性传播中可能发挥作用。测定了2.1 kb DNA转移区域的核苷酸序列。它在富含A+T的区域包含一个假定的转移起始点(oriT),包括三个反向重复序列和两个整合宿主因子结合位点。鉴定出的两个转移基因(mobA和mobB)组织在一个操纵子中,高效转移两者均是必需的。Southern印迹表明,质粒pIP417的转移区域与红霉素抗性质粒pBFTM1O和5-Ni抗性质粒pIP419的转移区域密切相关,但与5-Ni抗性质粒pIP421的转移区域无关。

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