Wallich R, Helmes C, Schaible U E, Lobet Y, Moter S E, Kramer M D, Simon M M
German Cancer Research Center, Heidelberg.
Infect Immun. 1992 Nov;60(11):4856-66. doi: 10.1128/iai.60.11.4856-4866.1992.
In order to assess the genetic variation of immunologically relevant structures among isolates of the Lyme disease spirochete, Borrelia burgdorferi, three chromosomal genes encoding flagellin (fla) and the heat shock proteins HSP60 and HSP70, as well as the plasmid gene encoding outer surface protein A (OspA), from 55 different European and North American strains obtained from ticks and mammal hosts have been investigated by restriction fragment length polymorphisms (RFLPs). RFLPs of fla and the HSP60 and HSP70 genes revealed two distinct banding patterns (A and B) for each of the three genes and allowed the definition of four genomic groups [AAA, BBB, BBA, and B(A/B)A] for the three chromosomal genes. On the other hand, RFLPs of the OspA gene revealed six distinct banding patterns (types I to VI) making up six independent genomic groups for the plasmid-encoded gene. Furthermore, we have sequenced the chromosomal HSP60 gene from B. burgdorferi ZS7 and the plasmid-encoded OspA gene from two strains, ZQ1 and 19857. Alignment of the deduced HSP60 amino acid sequence from B. burgdorferi ZS7 (genomic group AAA) to a previously published HSP60 sequence derived from strain ACA-1, which according to the proposed classification is in a different genomic group (BBA), revealed a sequence identity of > 99%. Similar alignments of the OspA sequence of strain ZQ1 to those of other isolates that were published previously revealed sequence identities of between 70 and 94% among strains of distinct OspA genomic groups. These data indicate the existence of a restricted number of species-specific subgroups and clearly show that genotypic variation is much more pronounced for the OspA gene than for fla and the HSP60 and HSP70 genes. A phylogenetic tree constructed on the basis of distance matrix analyses of 12 OspA sequences supports the proposed classification of genomic groups of B. burgdorferi.
为了评估莱姆病螺旋体伯氏疏螺旋体分离株中免疫相关结构的遗传变异情况，我们采用限制性片段长度多态性（RFLP）方法，对从蜱虫和哺乳动物宿主中获取的55株不同的欧洲和北美菌株的三个编码鞭毛蛋白（fla）、热休克蛋白HSP60和HSP70的染色体基因，以及编码外表面蛋白A（OspA）的质粒基因进行了研究。fla、HSP60和HSP70基因的RFLP分析显示，这三个基因各自呈现出两种不同的条带模式（A和B），据此可将三个染色体基因定义为四个基因组组[AAA、BBB、BBA和B(A/B)A]。另一方面，OspA基因的RFLP分析显示出六种不同的条带模式（I型至VI型），构成了该质粒编码基因的六个独立基因组组。此外，我们对伯氏疏螺旋体ZS7的染色体HSP60基因以及两个菌株ZQ1和19857的质粒编码OspA基因进行了测序。将伯氏疏螺旋体ZS7（基因组组AAA）推导的HSP60氨基酸序列与先前发表的源自ACA - 1菌株的HSP60序列进行比对，根据提议的分类，ACA - 1菌株属于不同的基因组组（BBA），结果显示序列同一性> 99%。将菌株ZQ1的OspA序列与先前发表的其他分离株的序列进行类似比对，结果显示不同OspA基因组组的菌株之间的序列同一性在70%至94%之间。这些数据表明存在数量有限的物种特异性亚组，并且清楚地表明OspA基因的基因型变异比fla、HSP60和HSP70基因更为明显。基于12个OspA序列的距离矩阵分析构建的系统发育树支持了伯氏疏螺旋体基因组组的提议分类。
