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由染色体编码的松弛酶和类TraG蛋白介导的幽门螺杆菌接合性质粒DNA转移

Conjugative plasmid DNA transfer in Helicobacter pylori mediated by chromosomally encoded relaxase and TraG-like proteins.

作者信息

Backert Steffen, Kwok Terry, König Wolfgang

机构信息

Department of Medical Microbiology, Otto von Guericke University, Leipziger Str. 44, D-39120 Magdeburg, Germany.

出版信息

Microbiology (Reading). 2005 Nov;151(Pt 11):3493-3503. doi: 10.1099/mic.0.28250-0.

Abstract

One of the striking characteristics of Helicobacter pylori is the extensive genetic diversity among clinical isolates. This diversity has been attributed to an elevated mutation rate, impaired DNA repair, DNA transfer and frequent recombination events. Plasmids have also been identified in H. pylori but it remained unknown whether conjugation can contribute to DNA transfer between clinical isolates. To examine whether H. pylori possesses intrinsic capability for conjugative plasmid transfer, shuttle vectors were introduced into H. pylori containing an oriT sequence of the conjugative IncPalpha plasmid RP4 but no mobilization (mob) genes. It was shown that these vectors could stably replicate and be mobilized among clinical H. pylori strains. It was also demonstrated that traG and relaxase (rlx) homologues carried on the H. pylori chromosome were important for plasmid transfer. Primer extension studies and mutagenesis further confirmed that the relaxase homologue rlx1 in H. pylori encodes a functional enzyme capable of acting on the RP4 oriT. Furthermore, the findings of this study indicate that traG and rlx1 act independently of the previously described type IV secretion systems, including that encoded by the cag pathogenicity island and the comB transformation apparatus, in mediating conjugative plasmid DNA transfer between H. pylori strains.

摘要

幽门螺杆菌的一个显著特征是临床分离株之间存在广泛的遗传多样性。这种多样性归因于突变率升高、DNA修复受损、DNA转移和频繁的重组事件。幽门螺杆菌中也已鉴定出质粒,但尚不清楚接合作用是否有助于临床分离株之间的DNA转移。为了研究幽门螺杆菌是否具有接合性质粒转移的内在能力,将穿梭载体导入含有接合性IncPα质粒RP4的oriT序列但无动员(mob)基因的幽门螺杆菌中。结果表明,这些载体能够在临床幽门螺杆菌菌株中稳定复制并被转移。还证明了幽门螺杆菌染色体上携带的traG和松弛酶(rlx)同源物对质粒转移很重要。引物延伸研究和诱变进一步证实,幽门螺杆菌中的松弛酶同源物rlx1编码一种能够作用于RP4 oriT的功能性酶。此外,本研究结果表明,traG和rlx1在介导幽门螺杆菌菌株之间的接合性质粒DNA转移时,独立于先前描述的IV型分泌系统发挥作用,包括由cag致病岛和comB转化装置编码的系统。

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