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Genetic Characteristics of Borrelia coriaceae Isolates from the Soft Tick Ornithodoros coriaceus (Acari: Argasidae)

机译:软壁OrOnithodoros coriaceus(Acari:Argasidae)的疏螺旋疏螺旋体的遗传特征

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摘要

Two Borrelia isolates (CA434 and CA435) cultured from the soft tick Ornithodoros coriaceus were analyzed by contour-clamped homogeneous electric field gel electrophoresis of unrestricted and ApaI-restricted DNA, standard electrophoresis of BamHI- and HindIII-restricted DNA, Southern hybridization, restriction fragment length polymorphism and sequencing of the 16S rRNA gene, and amplification of the 5S-23S intergenic spacer region. These isolates were compared with Borrelia coriaceae type strain Co53, B. burgdorferi sensu stricto strain CA4, and the relapsing-fever spirochete B. parkeri (undesignated). The 16S rRNA region of CA434 and CA435 differed from that of B. coriaceae type strain Co53 by the presence of 1 base (C) at position 367 (GenBank accession no. ). The linear plasmid profile of CA434 was similar to that of Co53, and the ApaI, BamHI, and HindIII restriction fingerprints of the total cellular DNA of CA434 and Co53 were similar. In contrast, CA435 differed somewhat from CA434 and Co53, which demonstrates that B. coriaceae is genetically diverse. Southern hybridization showed that the DNAs of CA434 and CA435 hybridized strongly with the digoxigenin-labeled DNA of Co53. Low homology was found between the DNA of Co53 and that of B. parkeri. The 16S rRNA sequence of B. parkeri was identical to previously published results for B. parkeri strain M3001 (GenBank accession number ). CA434 and CA435 represent only the second and third isolates of B. coriaceae obtained from any source since its initial isolation from an O. coriaceus tick in 1985. All three B. coriaceae isolates were derived from adult ticks collected from the same locality in northwestern California. Difficulties encountered in detecting B. coriaceae in, and isolating this spirochete from, the tissues of O. coriaceus are discussed. The lack of concordance between different detection or isolation methods suggests that reliance upon a single technique may grossly underestimate the true prevalence of spirochetal infection in wild-caught O. coriaceus ticks.
机译:通过轮廓限制的无限制和ApaI限制的DNA的均匀电场凝胶电泳,BamHI和HindIII限制的DNA的标准电泳,Southern杂交,限制片段,分析了从软壁虱Ornithodoros coriaceus培养的两种疏螺旋体(CA434和CA435)。长度多态性和16S rRNA基因的测序,以及5S-23S基因间隔区的扩增。将这些分离物与鲍氏疏螺旋体型菌株Co53,B.burgdorferi sensustricto菌株CA4和复发热螺旋体B.parkeri(未指定)进行了比较。 CA434和CA435的16S rRNA区域与B. coriaceae型菌株Co53的不同之处在于在367位(GenBank登录号)存在1个碱基(C)。 CA434的线性质粒图谱与Co53相似,并且CA434和Co53的总细胞DNA的ApaI,BamHI和HindIII限制性指纹图谱相似。相比之下,CA435与CA434和Co53有所不同,这证明了棒状杆菌属于遗传多样性。 Southern杂交表明,CA434和CA435的DNA与洋地黄毒苷标记的Co53的DNA强烈杂交。发现Co53的DNA和帕克氏杆菌的DNA之间的同源性低。帕克氏杆菌的16S rRNA序列与先前公布的帕克氏杆菌M3001菌株的结果相同(GenBank登录号)。 CA434和CA435仅代表 B的第二个和第三个分离株。自最初从 O分离出来以来,从任何来源获得的ia科。 coriaceus 在1985年打勾。所有三个 B。 ia科的分离株是从加利福尼亚西北部同一地区收集的成年tick中提取的。检测 B时遇到困难。菊科并从 O的组织中分离出这种螺旋体。讨论了coriaceus 。不同的检测或分离方法之间缺乏一致性,这表明对单一技术的依赖可能严重低估了野生 O中螺旋体感染的真实发生率。 coriaceus 滴答声。

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