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对莱姆病疏螺旋体中一个类VMP序列(vls)位点以及四种狭义伯氏疏螺旋体中的Vls同源物的分析。

Analysis of a VMP-like sequence (vls) locus in Borrelia garinii and Vls homologues among four Borrelia burgdorferi sensu lato species.

作者信息

Wang G, van Dam A P, Dankert J

机构信息

Department of Medical Microbiology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.

出版信息

FEMS Microbiol Lett. 2001 May 15;199(1):39-45. doi: 10.1111/j.1574-6968.2001.tb10648.x.

Abstract

The VMP-like sequence (vls) locus that consists of one expressed vlsE gene and 15 silent vls cassettes has been described in Borrelia burgdorferi sensu stricto B31. In the present study, the vls locus from a Borrelia garinii isolate A87SA was analyzed. DNA fragments that contained three complete and five partial vls cassettes were cloned and sequenced. Pulsed-field gel electrophoresis (PFGE) analysis and Southern hybridization of the PFGE blot indicated that the vls locus of B. garinii A87SA, consisting of at least eight vls cassettes, was located on a 21-kb linear plasmid. The size of the three complete vls cassettes varied from 573 to 612 bp. They had 93.8-94.3% identity at the nucleotide level and 84.9-87.3% amino acid identity. The amino acid sequences of the three vls cassettes of B. garinii A87SA exhibited 45.9-50.8% identity to the VlsE sequence of B. burgdorferi B31, and 30.0-33.8% identity to the VMP17 sequence of B. hermsii HS1. Homologues of the vls locus of B. garinii were detected by dot blot hybridization among 24 of the 30 (80.0%) isolates representing four B. burgdorferi sensu lato species distributed widely in Europe. Our findings indicate that B. garinii might possess a similar vls structure to that described in B. burgdorferi sensu stricto. The highly conserved nature of the vls locus among various B. burgdorferi sensu lato species suggests that it may be important in the physiology and pathogenesis of Lyme disease spirochetes.

摘要

在严格意义上的伯氏疏螺旋体B31中,已描述了由一个表达的vlsE基因和15个沉默的vls盒组成的类VMP序列(vls)位点。在本研究中,对一株加氏疏螺旋体分离株A87SA的vls位点进行了分析。克隆并测序了包含三个完整和五个部分vls盒的DNA片段。脉冲场凝胶电泳(PFGE)分析以及PFGE印迹的Southern杂交表明,加氏疏螺旋体A87SA的vls位点由至少八个vls盒组成,位于一个21 kb的线性质粒上。三个完整vls盒的大小在573至612 bp之间变化。它们在核苷酸水平上具有93.8 - 94.3%的同一性,在氨基酸水平上具有84.9 - 87.3%的同一性。加氏疏螺旋体A87SA的三个vls盒的氨基酸序列与伯氏疏螺旋体B31的VlsE序列具有45.9 - 50.8%的同一性,与赫氏疏螺旋体HS1的VMP17序列具有30.0 - 33.8%的同一性。通过斑点印迹杂交在代表广泛分布于欧洲的四种狭义伯氏疏螺旋体的30株分离株中的24株(80.0%)中检测到了加氏疏螺旋体vls位点的同源物。我们的研究结果表明,加氏疏螺旋体可能具有与严格意义上的伯氏疏螺旋体中所描述的类似的vls结构。vls位点在各种狭义伯氏疏螺旋体物种中的高度保守性质表明,它可能在莱姆病螺旋体的生理学和发病机制中很重要。

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