He Ping, Sheng Yue-Ying, Shi Yao-Zhou, Jiang Xiu-Gao, Qin Jin-Hong, Zhang Zhi-Ming, Zhao Guo-Ping, Guo Xiao-Kui
Department of Microbiology and Parasitology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
BMC Genomics. 2007 Jul 1;8:204. doi: 10.1186/1471-2164-8-204.
Leptospirosis is a world-widely distributed zoonosis. Humans become infected via exposure to pathogenic Leptospira spp. from contaminated water or soil. The availability of genomic sequences of Leptospira interrogans serovar Lai and serovar Copenhageni opened up opportunities to identify genetic diversity among different pathogenic strains of L. interrogans representing various kinds of serotypes (serogroups and serovars).
Comparative genomic hybridization (CGH) analysis was used to compare the gene content of L. interrogans serovar Lai strain Lai with that of other 10 L. interrogans strains prevailed in China and one identified from Brazil using a microarray spotted with 3,528 protein coding sequences (CDSs) of strain Lai. The cutoff ratio of sample/reference (S/R) hybridization for detecting the absence of genes from one tested strain was set by comparing the ratio of S/R hybridization and the in silico sequence similarities of strain Lai and serovar Copenhageni strain Fiocruz L1-130. Among the 11 strains tested, 275 CDSs were found absent from at least one strain. The common backbone of the L. interrogans genome was estimated to contain about 2,917 CDSs. The genes encoding fundamental cellular functions such as translation, energy production and conversion were conserved. While strain-specific genes include those that encode proteins related to either cell surface structures or carbohydrate transport and metabolism. We also found two genomic islands (GIs) in strain Lai containing genes divergently absent in other strains. Because genes encoding proteins with potential pathogenic functions are located within GIs, these elements might contribute to the variations in disease manifestation. Differences in genes involved in O-antigen biosynthesis were also identified for strains belonging to different serogroups, which offers an opportunity for future development of genomic typing tools for serological classification.
CGH analyses for pathogenic leptospiral strains prevailed in China against the L. interrogans serovar Lai strain Lai CDS-spotted microarrays revealed 2,917 common backbone CDSs and strain specific genes encoding proteins mainly related to cell surface structures and carbohydrated transport/metabolism. Of the 275 CDSs considered absent from at least one of the L. interrogans strains tested, most of them were clustered in the rfb gene cluster and two putative genomic islands (GI A and B) in strain Lai. The strain-specific genes detected via this work will provide a knowledge base for further investigating the pathogenesis of L interrogans and/or for the development of effective vaccines and/or diagnostic tools.
钩端螺旋体病是一种广泛分布于全球的人畜共患病。人类通过接触受污染水或土壤中的致病性钩端螺旋体而感染。问号钩端螺旋体赖型和哥本哈根型的基因组序列的可得性为鉴定代表各种血清型(血清群和血清型)的问号钩端螺旋体不同致病菌株间的遗传多样性提供了机会。
利用比较基因组杂交(CGH)分析,通过用点有问号钩端螺旋体赖型菌株赖型的3528个蛋白质编码序列(CDS)的微阵列,比较了问号钩端螺旋体赖型菌株赖型与中国流行并从巴西鉴定出的其他10株问号钩端螺旋体的基因含量。通过比较赖型菌株与问号钩端螺旋体哥本哈根型菌株Fiocruz L1 - 130的S/R杂交比率和电子序列相似性,设定检测受试菌株中基因缺失的样本/参照(S/R)杂交的截止比率。在所检测的11株菌株中,发现至少有一株菌株缺失275个CDS。问号钩端螺旋体基因组的共同主干估计包含约2917个CDS。编码诸如翻译、能量产生和转换等基本细胞功能的基因是保守的。而菌株特异性基因包括那些编码与细胞表面结构或碳水化合物运输及代谢相关蛋白质的基因。我们还在赖型菌株中发现了两个基因组岛(GI),其中含有的基因在其他菌株中明显缺失。由于编码具有潜在致病功能蛋白质的基因位于基因组岛内,这些元件可能导致疾病表现的差异。还鉴定出属于不同血清群的菌株在O抗原生物合成相关基因上的差异,这为血清学分类的基因组分型工具的未来开发提供了机会。
针对问号钩端螺旋体赖型菌株赖型CDS点样微阵列,对中国流行的致病性钩端螺旋体菌株进行的CGH分析揭示了2917个共同主干CDS以及编码主要与细胞表面结构和碳水化合物运输/代谢相关蛋白质的菌株特异性基因。在所检测的问号钩端螺旋体菌株中,至少有一株菌株缺失的275个CDS中,大多数聚集在rfb基因簇以及赖型菌株中的两个假定基因组岛(GI A和B)中。通过这项工作检测到的菌株特异性基因将为进一步研究问号钩端螺旋体的发病机制和/或开发有效的疫苗和/或诊断工具提供知识库。