Correlation between plasmid content and infectivity in Borrelia burgdorferi
Correlation between plasmid content and infectivity in Borrelia burgdorferi
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DOI：
10.1073/pnas.97.25.13865
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发表时间：
2000-12-05
影响因子：
11.1
通讯作者：
Norris, SJ
中科院分区：
文献类型：
--
作者：
Purser, JE;Norris, SJ
通过使用PCR检测一组19个克隆分离株中的每一个质粒，在伯氏疏螺旋体B31中鉴定出与繁殖力相关的质粒。克隆表现出高，低，和中间感染性表型的基础上，他们的频率从针接种C3 H/HeN小鼠分离。通过使用对每个质粒特有的区域特异的PCR引物，确定22个质粒中的21个在每个克隆中的存在或不存在，如在最近可获得的基因组序列中鉴定的。Southern杂交结果用于证实部分病例的PCR结果。质粒lp 25表现出与感染性的直接相关性，因为它始终存在于所有高或中等感染性的克隆中，而在所有低感染性的克隆中不存在。含有VP样序列基因座的LP 28 -1也与感染性相关;所有缺乏LP 28 -1但含有LP 25的克隆都具有中等感染性表型，其中感染主要限于关节。质粒cp 9，cp 32 -3，lp 21。在该模型中，lp 28 -2、lp 28 -4和lp 56显然不是感染所必需的，因为缺乏这些质粒的克隆表现出高感染性表型。质粒cp 26，cp 32 -1。cp 32 -2和/或cp 32 -7、cp 32 -4、cp 32 -6、cp 32 -8、cp 32 -9、lp 17、lp 28 -3、lp 36、lp 38和lp 54一致地存在于所检测的所有克隆中。基于这些结果，lp 25和lp 28 -1似乎编码在B的发病机制中重要的毒力因子。Burgdorferi B31.
Infectivity-associated plasmids were identified in Borrelia burgdorferi B31 by using PCR to detect each of the plasmids in a panel of 19 clonal isolates. The clones exhibited high-, low-, and intermediate-infectivity phenotypes based on their frequency of isolation from needle-inoculated C3H/HeN mice. presence or absence of 21 of the 22 plasmids was determined in each of the clones by using PCR primers specific for regions unique to each plasmid, as identified in the recently available genome sequence. Southern blot hybridization results were used to confirm the PCR results in some cases. Plasmid lp25 exhibited a direct correlation with infectivity in that it was consistently present in all clones of high or intermediate infectivity and was absent in all low-infectivity clones. lp28-1, containing the vmp-like sequence locus, also correlated with infectivity; all clones that lacked lp28-1 but contained lp25 had an intermediate infectivity phenotype, in which infection was primarily restricted to the joints. Plasmids cp9, cp32-3, lp21. lp28-2, lp28-4, and lp56 apparently are not required for infection in this model, because clones lacking these plasmids exhibited a high-infectivity phenotype. Plasmids cp26, cp32-1. cp32-2 and/or cp32-7, cp32-4, cp32-6, cp32-8, cp32-9, lp17, lp28-3, lp36, lp38, and lp54 were consistently present in ail clones examined. On the basis of these results, lp25 and lp28-1 appear to encode virulence factors important in the pathogenesis of B. burgdorferi B31.