抑制差减杂交克隆E1 Tor霍乱弧菌流行株与非流行株基因组差异片段及其分析  

Identification of Genomic Differential Fragments Between Epidemic and Nonepidemic Strains of Vibrio choleraeO1 E1 Tor by Using SSH

作　　者：张丽娟[1] 阚飙[1] 杨帆[2] 高守一[1] 刘延清[1] 祁国明[1] 

机构地区：[1]中国预防医学科学院流行病学微生物学研究所卫生部医学分子细菌学重点实验室,北京102206 [2]中国预防医学科学院病毒基因工程国家重点实验室,北京100052

出　　处：《中国生物化学与分子生物学报》2002年第5期573-577,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究"973"计划资助基金项目 (No .G19990 5 410 2 )~~

摘　　要：为了克隆与分析霍乱弧菌O1E1Tor流行株与非流行株两类菌株基因组差异片段 ,采用抑制差减杂交技术 (suppressionsubtractivehybridization ,SSH)分别以国内保留的流行株Wujiang 2及非流行株Js 32一株作为被检菌 ,另一株作为参考菌进行基因组差异研究 .在进行的差减杂交实验中 ,流行株Wujiang 2共检出 34个特异差异片段 ,经同源检索共代表 35个基因片段 ,其中包括许多重要的霍乱弧菌毒力相关基因如CTX遗传单元、TLC因子及可移动外来成分如霍乱弧菌整合子RVC序列及Tn10转位酶 .非流行株Js 32共检出 14个特异差异片段 ,经同源检索未能检出同源片段 ,可能代表新基因序列 .研究表明 ,霍乱弧菌流行株与非流行株基因组存在较多差异基因 ,表现在毒力、毒力相关基因。In order to study the genetic differences between epidemic and nonepidemic strains of Vibrio choleraeO1 E1 Tor, the suppression subtractive hybridization was used to screen for the different genes between epidemic and nonepidemic strains of V. cholerae. Strain Wujiang-2 is defined as epidemic strain whereas Js-32 is defined as nonepidemic strain on the basis of their phage-biotypes. 34 fragments specific to the epidemic strain of Wujiang-2 representing 35 genes were obtained by SSH against the nonepidemic strain of Js-32. On the other hand, 14 fragments specific to the nonepidemic strain of Js-32 identified by SSH, were present in nonepidemic strain and absent in the other. The results suggest that there exist some genetic differences between epidemic and nonepidemic strains of V. cholerae.

关 键 词：抑制差减杂交 EL Tor霍乱弧菌流行株 非流行株 基因组差异片段 序列分析 基因克隆 

分 类 号：R378.3[医药卫生—病原生物学] R516.5[医药卫生—基础医学]