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Cloning and sequence analysis of Helicobacter pylori ureC

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Author:: No author available

Journal Title:: WORLD CHINESE JOURNAL OF DIGESTOLOGY

Issue:: 3

DOI:: 10.3969/j.issn.1009-3079.2001.03.014

Key Word:: 尿素酶/遗传学;螺杆菌,幽门/酶学;螺杆菌,幽门/分离和提纯;聚合酶链反应

Abstract：目的用PCR方法扩增幽门螺杆菌(Hp)尿素酶C基因,以构建Hp UreC基因的重组克隆,为进一步表达Hp UreC基因的重组蛋白质并研究其功能奠定基础.方法以Hp临床菌株总DNA为模板,根据已发表的HpUreC基因序列设计引物(位于588bp～605bp和1737bp～1754bp),采用PCR方法扩增出一个1173bp的Hp尿素酶C基因片段,用BanHI及Eco R I双酶切后将其克隆入测序质粒pGEM-3Zf(-)中,以全自动测序仪双向测定目的片段序列,借助于DNA TOOL 5.1拼接成完整序列,与已知的Hp UreC基因序列做比较.结果所得核酸序列与报道的Hp国际标准菌株M60398的UreC基因同源性为95%,根据测序结果推断其编码的氨基酸序列,并行BLAST分析,发现它与标准菌株M60398的氨基酸序列同源性为97%结论成功构建了Hp UreC基因的重组克隆,为进一步研究表达Hp UreC基因的重组蛋白质并研究其功能奠定了基础.

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