期刊论文

Li, C.

中文

微生物学报

0001-6209

2008

48

3

349-354

10.3321/j.issn:0001-6209.2008.03.013

湖南省自然科学基金（06JJ2093） 湖南省教育厅资助项目（05C471）

本校为第一机构

构建pDsRed-Monomer-C1／vacA N端真核表达载体，研究幽门螺杆菌空泡毒素单一毒力决定簇对THP-1巨噬细胞细胞因子分泌的影响。PCR扩增vacA目的基因片段，克隆人真核表达载体pDsRed—Monomer-C1中，经酶切、PCR及测序鉴定后，转染THP-1巨噬细胞中，Western blot和荧光显微镜鉴定VacA蛋白在细胞中的表达；电子显微镜和中性红摄人法观察巨噬细胞的空泡样变；ELISA法检测巨噬细胞培养上清TNF-α、IL-1β含量。重组质粒转染THP-1巨噬细胞24h，部分细胞胞浆中出现大小不等的空泡，且重组质粒组培养上清中TNF-α、IL-1β含量明显高于空质粒组和阴性对照组（P〈0．001），二硫代氨基甲酸吡咯烷（pyrroli...

We constructed a recombinant plasmid containing the N-terminus gene of vacA gene of Helicobacter pylori and studied the effect of VacA on the secretion of macrophages as an individual virulence determinant. VacA gene amplified by Polymerase Chain Reaction (PCR) from Helicobacter pylori was cloned into eukaryotic expression vector pDsRed-Monomer-C1. The recombinant plasmids were verified by restriction endonucleases analysis and nucleotide sequencing. Then the recombinant plasmids pDsRed-Monomer-C1/vacA were transfected into macrophages. Their expression in macrophages was examined by Western b...