中国兽医科学2013，Vol.43Issue(3)：283-288,6.
幽门螺杆菌尿素酶B基因的克隆表达及活性测定
Cloning, expression and functional analysis of the UreB gene from Helicobacter pylori
蔡靓 1李克生 2杜惠芬 3付宝权 2李文卉 2曲自刚 3谢志宙3
作者信息
- 1. 甘肃农业大学动物医学院,甘肃兰州730070
- 2. 甘肃省医学科学研究院医学生物中心,甘肃兰州 730050
- 3. 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部兽医公共卫生重点开放实验室甘肃省动物寄生虫病重点实验室,甘肃兰州 730046
- 折叠
摘要
Abstract
In order to study the immunological activities of urease B subunit(UreB) from Helicobacter pylori(Hp) ,UreB gene of Hp was amplified by PCR,digested with restriction enzyme and cloned into the plasmid pET-28a. The recombinant expression vector pET28a-UreB was transformed into Escherichia coli BL2KDE3) strains,and recombinant protein UreB was expressed by IPTG induction. BALB/c mice were immunized with purified recombinant UreB protein,and antibody titer was measured by ELISA. The immu-noreactivity of recombinant protein was identified by Western-blot. The results indicated that the homology of nucleotide sequence of the cloned UreB gene was not less than 96. 73% in comparison with 26 695 standard strain sequences in GenBank, while the homology of its putative amino acid sequence was 99. 30%. SDS-PAGE analysis showed that the molecular weight of recombinant protein was about 66 ku and presented in the supernatant. The purified recombinant protein could react with serum from patients infected by Hp in Western-blot,specific humoral immune response could be induced by purified protein,which showed that the recombinant protein had immunogenicity. The results showed that the recombinant protein UreB was expressed successfully and possessed strong immunogenicity,which will be contributed to the develop-
merit of rapid detection methods for detection of Hp infection.关键词
幽门螺杆菌/尿素酶B亚单位/原核表达/免疫原性Key words
Helicobacter pylori/UreB subunit/prokaryotic expression/immunogenicity分类
农业科技引用本文复制引用
蔡靓,李克生,杜惠芬,付宝权,李文卉,曲自刚,谢志宙..幽门螺杆菌尿素酶B基因的克隆表达及活性测定[J].中国兽医科学,2013,43(3):283-288,6.
