Comprehensive analyses of bacterial pathogenesis based on the genome information

本研究基于三种重要病原菌的基因组序列，对它们的致病性和基因组多样性进行了综合分析，主要研究结果如下：1.致病性大肠杆菌：（1）基于O 157 Sakai基因序列，建立了基因芯片和全基因组PCR扫描方法。(2)利用这两种方法，我们对O 157和非O 157肠出血性大肠杆菌进行了大规模的基因组比较。结果表明，O 157菌株间存在显著的基因组多样性，原噬菌体和IS元件的变异是造成这种基因组多样性的主要因素。此外，我们发现，非O 157 EHEC菌株含有大量的DNA，不存在于O 157。(3)我们发现RcsCDB双组分调控系统和碳酸氢盐敏感系统控制着毒力基因的表达，并发现了新的调控因子GrvA和Pch，它们参与了这些调控。 关于我们 离子系统通过表达谱分析使用微阵列。这为了解致病性大肠杆菌的毒力基因表达系统是如何调控的提供了线索。大肠杆菌的内在调控网络。杆菌(4)通过采用蛋白质组学方法，我们发现了O157.2的III型分泌系统（TTSS）的许多新效应子。产气荚膜梭菌：（1）测定了13号菌株的基因组序列。(2)利用基于其基因组序列构建的基因芯片，我们鉴定了141个受VirR/S双组分调控系统或VirR/S-VR-RNA系统调控的基因。它们不仅包括毒力基因，而且还包括许多与能量产生和代谢有关的基因，表明毒力基因和生长基因在宿主组织中受到VirR/S-VR-RNA系统的协调调控。(3)通过对VirR结合序列的搜索，我们发现了两个新的调控RNA.（1）测定了副溶血性弧菌RIMD 2210633菌株的基因组序列。(2)我们在其基因组上发现了两个TTSS（TTSS 1和TTSS 2），并发现TTSS 1与细胞毒性有关，TTSS 2与致瘤性有关。(3)我们为每个TTSS确定了几个效应子，并发现这两个TTSS表现出不同的效应子特异性。少

In this research, we performed a comprehensive analysis of the pathogenesis and genomic diversity of three important pathogenic bacteria based on their genome sequences, and obtained followings major findings.1. Pathogenic Escherichia coli:(1) Based on the O157 Sakai sequence, we developed ○a microarray and the whole genome PCR scanning method. (2) By using these two methods, we performed a large-scale genomic comparison of O157 and non-O157 enterohemorrhagic E. coli (EHEC) strains, and revealed that there exits a remarkable genomic diversity among O157 strains, and that variation of prophages and IS elements are major factors to generate this genomic diversity. Furthermore, we revealed that non-O157 EHEC strains contain a large amount of DNAs that are not present in O157. (3) We found that the RcsCDB two component regulatory system and the bicarbonate-sensing system control the virulence gene expression, and discovered novel regulators, GrvA and Pch, that are involved in these regulat … More ion systems by expression profiling anakyses using the microarray. This provided the first clue to understand how the virulence gene expression systems of pathogenic E. coli are incorporated into the intrinsic regulatory network of E. coli. (4) By employing a proteomic approach, we discovered many new effectors for the type III secretion system (TTSS) of O157.2. Clostridium perfringen :(1) The genome sequence of strain 13 was determined. (2) By using the microarray constructed based on its genome sequence, we identified 141 genes that are controlled by the VirR/S two component regulatory system or the VirR/S-VR-RNA system. They included not only virulence genes but also many genes for energy production and metabolism, indicating that genes for virulence and growth in host tissues are coordinately regulated by the VirR/S-VR-RNA system. (3) By searching the VirR-binding sequences, we identified two novel regulatory RNAs.3. Vibrio parahaemolyticus :(1) The genome sequence of strain RIMD2210633 was determined. (2) We identified two TTSSs (TTSS1 and TTSS2) on its genome, and found that TTSS1 is involved in cell toxicity and TTSS2 in diarrheagenicity. (3) We identified several effectors for each TTSS, and found that the two TTSS s exhibit distinct effector-specificities. Less