【摘要】 目的获取人临床株幽门螺旋杆菌(Hp)热休克蛋白A(HspA)亚单位基因。方法用聚合酶链反应(PCR)技术从Hp的染色体DNA扩增HspA基因,将其定向插入pMD18-T克隆载体中进行核苷酸序列分析。结果经酶切鉴定及基因序列证实Hp HspA基因克隆成功。克隆载体pMD-HspA测序结果显示HspA DNA片段长度为357 bp。利用生物软件Om iga 2.0对临床分离的Hp和NCTC11637 HspA基因序列进行同源性比较,发现有14个核苷酸差异,基因序列同源性96.08%;对编码氨基酸进行比较,发现有1个氨基酸差异,氨基酸同源性99.15%。同时将Hp HspA序列与GenBank中公布的多株国外Hp HspA序列进行比较,结果显示基因序列同源性在95.20%~96.36%之间,氨基酸序列同源性在98.31%~100%之间。结论成功地获得人临床株Hp保守的HspA基因,为Hp HspA相关疫苗的研究奠定了实验基础。更多还原

【Abstract】 Objective To obtain the heat shock protein A(HspA) subunit gene in human clinical strains helicobacter pylori(Hp).Methods The HspA gene form chromosome DNA of Hp was amplified by polymerase chain reaction(PCR) technique,and the obtained DNA fragment were inserted into pMD18-T cloning vector for nucleotide sequence analysis.Results Hp HspA gene was cloned successfully,which was demonstrated by enzyme digesting and gene sequencing.Sequencing result of cloning vector pMD-HspA showed that the length of HspA DNA fragment was 357 bp.HspA gene sequence homology between Hp separated form clinic and NCTC11637 was compared by biological software Omiga 2.0 and 14 nucleotide differences were found.Gene sequence homology was 96.08%;Compared with coded amino acids,1 amino acids difference was found.Amino acids homology was 99.15%.At the same time,when compared with Hp HspA sequence reported in Genbank,the result showed that gene sequence homology was 95.20%-96.36%,and amino acids sequence homology was 98.31%-100%.Conclusion Conservative HspA gene in human clinical strains Hp was obtained successfully,which provided an experimental foundation for Hp HspA vaccine research.更多还原