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来自复发性发热病原体土拉疏螺旋体的一个多拷贝、线性质粒携带、含重复基序基因的克隆及分子特征分析

Cloning and molecular characterization of a multicopy, linear plasmid-carried, repeat motif-containing gene from Borrelia turicatae, a causative agent of relapsing fever.

作者信息

Carlyon J A, Marconi R T

机构信息

Department of Microbiology and Immunology, Medical College of Virginia at Virginia Commonwealth University, Richmond, Virginia 23298-0678, USA.

出版信息

J Bacteriol. 1998 Sep;180(18):4974-81. doi: 10.1128/JB.180.18.4974-4981.1998.

Abstract

Borrelia turicatae is one of several spirochete species that can cause relapsing fever. Here, we describe the identification and characterization of a gene from B. turicatae and other relapsing-fever spirochetes that exhibits homology with the rep+ and ORF-E gene families of the Lyme disease spirochetes. This gene, which we have designated repA, encodes a putative protein of 30.2 kDa with an isoelectric point of 4.69. The central region of RepA harbors a series of amino acid repeat motifs which exhibit homology with casein kinase 2 phosphorylation sites. Through Southern hybridization analyses, we demonstrate that repA (or a closely related sequence) is multicopy in the relapsing-fever spirochetes and is carried on variably sized linear plasmids in both Borrelia parkeri and B. turicatae. Transcriptional analyses demonstrate that repA is expressed, albeit at low levels, during in vitro cultivation of B. turicatae. Transcriptional start site analysis revealed that repA is preceded by a consensus ribosomal binding site and an appropriately spaced promoter element. The sequence conservation, unique features, and multicopy status of repA and its homologs suggest that RepA may play an important genus-wide role in the biology of the Borrelia.

摘要

杜氏疏螺旋体是几种可引起回归热的螺旋体物种之一。在此,我们描述了来自杜氏疏螺旋体和其他回归热螺旋体的一个基因的鉴定和特征,该基因与莱姆病螺旋体的rep +和ORF - E基因家族具有同源性。我们将该基因命名为repA,它编码一个推定的蛋白质,分子量为30.2 kDa,等电点为4.69。RepA的中央区域含有一系列氨基酸重复基序,与酪蛋白激酶2磷酸化位点具有同源性。通过Southern杂交分析,我们证明repA(或一个密切相关的序列)在回归热螺旋体中是多拷贝的,并且在帕克疏螺旋体和杜氏疏螺旋体中都存在于大小可变的线性质粒上。转录分析表明,在杜氏疏螺旋体的体外培养过程中,repA虽表达水平较低,但仍有表达。转录起始位点分析显示,repA之前有一个共有核糖体结合位点和一个间隔适当的启动子元件。repA及其同源物的序列保守性、独特特征和多拷贝状态表明,RepA可能在疏螺旋体生物学中发挥重要的全属范围作用。

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