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Borrelia burgdorferi Surface-Localized Proteins Expressed during Persistent Murine Infection Are Conserved among Diverse Borrelia spp.▿ †

机译:持久性鼠感染过程中表达的疏螺旋体表面固定蛋白在不同的疏螺旋体中均得到保留。

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摘要

Borrelia burgdorferi, the causative agent of Lyme disease in the United States, regulates numerous genes encoding lipoproteins on linear plasmid 54 in response to environmental cues. We analyzed a subset of these genes/proteins that were historically categorized as paralogous gene family 54 (BBA64, BBA65, BBA66, BBA68, BBA69, BBA70, BBA71, and BBA73) and found that the expression of several genes was influenced by the σN-σS regulatory cascade at the level of transcription and protein synthesis. Moreover, we established in this and a previous study that BBA65, BBA66, BBA69, BBA71, and BBA73 are temporally expressed during persistent infection of immunocompetent mice, as determined by quantitative real time-PCR of ear tissue, by enzyme-linked immunosorbent assay, and by immunoblotting. Correspondingly, BBA65, BBA66, BBA71, and BBA73 proteins were detectable in infectious B. burgdorferi B31 isolates but undetectable in noninfectious isolates. BBA65, BBA66, BBA71, and BBA73 proteins were also found to partition into the Triton X-114 detergent phase and were sensitive to protease treatment of intact cells, indicating that they are membrane associated and surface localized. Lastly, Southern blotting and PCR with specific gene primer/probes for BBA64, BBA65, BBA66, BBA71, and BBA73 suggest that many of these genes are conserved among the B. burgdorferi sensu lato isolates and the relapsing-fever Borrelia species. Together, the data presented suggest that these genes may play a part in Borrelia infection and/or pathogenicity that could extend beyond the sensu lato group.
机译:伯氏疏螺旋体(Borrelia burgdorferi)是美国莱姆病的病原体,可根据环境因素调节线性质粒54上编码脂蛋白的许多基因。我们分析了这些基因/蛋白质的一个子集,这些子集历史上被归类为旁系基因家族54(BBA64,BBA65,BBA66,BBA68,BBA69,BBA70,BBA71和BBA73),并发现几个基因的表达受σN-的影响。 σS在转录和蛋白质合成水平上具有调控级联作用。此外,我们在这项研究和先前的研究中确定,通过免疫组织化学定量耳部PCR(实时定量PCR)测定的,有免疫能力的小鼠持续感染过程中,BBA65,BBA66,BBA69,BBA71和BBA73会暂时表达,并通过免疫印迹。相应地,在感染性伯氏疏螺旋体B31分离物中可检测到BBA65,BBA66,BBA71和BBA73蛋白,而在非感染性分离物中则不可检测到。还发现BBA65,BBA66,BBA71和BBA73蛋白可以分配到Triton X-114去污剂相中,并且对完整细胞的蛋白酶处理敏感,表明它们与膜相关并且位于表面。最后,针对BBA64,BBA65,BBA66,BBA71和BBA73的特异性基因引物/探针的Southern杂交和PCR表明,其中许多基因在B. burgdorferi sensu lato分离株和复发性发热的Borrelia菌种中都是保守的。在一起,所提供的数据表明这些基因可能在疏螺旋体的感染和/或致病性中发挥作用,而这种影响可能会扩展到超感官组之外。

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