Cadavid D, Pennington P M, Kerentseva T A, Bergström S, Barbour A G
Department of Microbiology, University of Texas Health Science Center at San Antonio, 78284, USA.
Infect Immun. 1997 Aug;65(8):3352-60. doi: 10.1128/iai.65.8.3352-3360.1997.
In mice infected with serotype A but not serotype B of the relapsing fever spirochete Borrelia turicatae, early invasion of the brain occurs. Serotypes A and B are further distinguished by the abundant surface protein they produce: VmpA and VmpB, respectively. Western blotting with monoclonal antibodies, one-dimensional peptide mapping, and partial amino acid sequencing demonstrated regions of the VmpA protein that differed from VmpB. Oligonucleotide primers based on the partial amino acid sequences of unique regions were used to amplify a portion of the VmpA gene (vmpA) by PCR, and the product was used as a probe in Southern blot and Northern blot analyses. These experiments showed that (i) expression of the vmpA sequence was determined at the level of transcription and (ii) the vmpA sequence was in two locations in serotype A and one location in serotype B. The vmpA gene at the expression-linked locus of serotype A was cloned and sequenced. An open reading frame would encode a polypeptide of 214 amino acids. The polypeptide expressed by Escherichia coli was bound by VmA-specific but not VmpB-specific antibody. Primer extension analysis identified a consensus sigma70-type promoter for vmpA at the expression locus. Phylogenetic analysis revealed that VmpA is homologous to small Vmp (Vsp) proteins of B. hermsii and to OspC proteins of B. burgdorferi. These findings indicate that a function of the Vsp-OspC family of proteins of Borrelia spp. may be differential localization in organs, including the brain, during infection.
在感染了回归热螺旋体杜氏疏螺旋体血清型A而非血清型B的小鼠中,会出现早期脑部感染。血清型A和B的进一步区别在于它们产生的丰富表面蛋白:分别为VmpA和VmpB。用单克隆抗体进行蛋白质印迹分析、一维肽图分析以及部分氨基酸测序,结果显示VmpA蛋白与VmpB不同的区域。基于独特区域的部分氨基酸序列设计的寡核苷酸引物,用于通过聚合酶链反应(PCR)扩增VmpA基因(vmpA)的一部分,该产物用作Southern印迹和Northern印迹分析的探针。这些实验表明:(i)vmpA序列的表达是在转录水平上决定的;(ii)vmpA序列在血清型A中有两个位置,在血清型B中有一个位置。血清型A表达相关位点的vmpA基因被克隆并测序。一个开放阅读框将编码一个含214个氨基酸的多肽。大肠杆菌表达的该多肽能与VmA特异性抗体结合,而不能与VmpB特异性抗体结合。引物延伸分析在表达位点鉴定出vmpA的一个共有sigma70型启动子。系统发育分析表明,VmpA与赫氏疏螺旋体的小Vmp(Vsp)蛋白以及伯氏疏螺旋体的OspC蛋白同源。这些发现表明,疏螺旋体属的Vsp - OspC家族蛋白在感染过程中的一个功能可能是在包括大脑在内的器官中进行差异定位。