幽门螺杆菌VacA真核表达载体的构建及其诱导细胞空泡化和凋亡  

HELICOBACTER PYLORI VACA INDUCES VACUOLE FORMATION AND APOPTOSIS BY TRANSFECTION OF EUKARYOTIC EXPRESSION VECTOR OF VACA GENE

作　　者：袁建平[1] 赵蔚[1] 杨贵珍[1] 胡宝瑜[1] 童善庆[1] 郭晓奎[1] 

机构地区：[1]上海第二医科大学病原生物学教研室,上海200025

出　　处：《中国微生态学杂志》2003年第2期66-68,共3页Chinese Journal of Microecology

基　　金：国家教委留学归国基金

摘　　要：目的 :研究幽门螺杆菌空泡毒素作为单一毒力决定簇对真核细胞的作用。方法 :用 PCR扩增vac A基因片段 ,克隆入真核表达载体 p EGFP- N 1,构建重组质粒 p EGFP- vac A,转染 He L a细胞 ,通过相差显微镜和电子显微镜观察细胞形态与结构的变化。结果 :重组质粒转染 He L a细胞 2 4 h,10 %～ 2 0 %细胞的胞质内出现明显空泡 ,其中少数细胞发生凋亡改变。结论 :成功构建了用于真核表达的重组 vac A质粒 ,转染真核细胞后 ,观察 Vac A作用导致的细胞形态结构的变化 ,为研究 VacObjective:To investigate the role of VacA plays in eukaryocyte acting as an individual virulence determinant Methods:VacA gene of Helicobacter pylori amplified by PCR was cloned into eukaryotic expression vector pEGFP N1,resulting in a recombination plasmid pEGFP vacA HeLa cells were transfected with the plasmid encoding VacA,and then was detected by contrast microscopy and electron microscopy Results:24 h after the transfection,a clear vacuolated phenotype developed in 10%～20% of cells,and apoptosis arose in a minority of them Conclusion:The eukaryotic expression plasmid encoding VacA is successfully constructed,and the morphological alteration is detected after transfecting the plasmid into HeLa cells,providing a basis for investigating the role of VacA plays in eukaryocyte acting as an individual virulence determinant

关 键 词：幽门螺杆菌 空泡毒素 单一毒力决定簇 真核表达载体 细胞空泡化 细胞凋亡 

分 类 号：R378.99[医药卫生—病原生物学]