幽门螺杆菌5种候选疫苗抗原基因的克隆、表达及抗原性的鉴定
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宁云山, 李 妍, 龙 敏, 董文其, 李 明
幽门螺杆菌;候选疫苗抗原;克隆;基因表达;抗原性宁云山,李妍,龙敏,董文其,李明.幽门螺杆菌5种候选疫苗抗原基因的克隆、表达及抗原性的鉴定. 世界华人消化杂志2006;14(26)2605
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宁云山, 李妍, 龙敏, 董文其, 李明, 南方医科大学生物技术学院 广东省广州市 510515
宁云山, 2001年第一军医大学博士, 副教授, 主要从事蛋白质与抗体工程研究.
通讯作者: 李明, 510515, 广东省广州市同和, 南方医科大学生物技术学院. mingli@fimmu.com
电话: 020-61648550 传真: 020-61648550
收稿日期: 2006-03-08 接受日期: 2006-05-24
Cloning, expression and antigenicity identification of five candidate vaccine antigen genes of human Helicobacter pylori
Yun-Shan Ning, Yan Li, Min Long, Wen-Qi Dong, Ming Li
Yun-Shan Ning, Yan Li, Min Long, Wen-Qi Dong, Ming Li, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong Province, China
Correspondence to: Dr. Ming li, School of Biotechnology, Southern Medical University, Tonghe, Guangzhou 510515, Guangdong Province, China. mingli@fimmu.com
Received: 2006-03-08 Accepted: 2006-05-24
AbstractAIM:To construct a recombinant plasmid contai-ning genes encoding Lpp20, HspA, UreaseA, CagA, UreaseB from H pylori, express it in E.coli and explore the antigenicity.
METHODS:The genes, encoding Lpp20, HspA, UreaseA, CagA, and UreaseB, were amplified from H pylori chromosomal DNA by polymerase chain reaction (PCR), and then T-A was cloned and sequenced. The target genes cloned into pGEX-4T-1 fusion expression vector were expressed in E.coli and purified by GST-affinity chromatography. The purified products were used to identify 29 strains of mouse monoclonal antibodies (mAbs) against H pylori, and the antigenicity of the products was analyzed by Western blot with serum ofH pylori-infected patients.
RESULTS:Lpp20, HspA, UreaseA, CagA, and UreaseB fragments were 528 bp, 351 bp, 675 bp, 855 bp, and 1704 bp in length, respectively (GenBank submission No. DQ106902, DQ141574, DQ141577, DQ141575, and DQ141576, respectively), and the nucleotide homology was 95%-99% with other H pylori strains. Lpp20, HspA, UreaseA, CagA, and UreaseB fusion protein were expressed with molecular weights of 48 000, 41 000, 52 000, 6 0000, and 91 000 Da in E ......
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