人幽门螺杆菌HspA亚单位的基因克隆及序列分析

Cloning and Sequence Analysis of Human Helicobacter pylori HspA Subunit Gene

目的:获取幽门螺杆菌(Hp)热休克蛋白A(HspA)亚单位基因并进行核苷酸序列分析和同源性比较。方法:获取幽门螺杆菌的染色体DNA,PCR技术扩增获取HspA基因,将其定向插入pGEX-4T-1原核表达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HpHspA基因克隆成功,DNA序列与Genbank公布的序列比较有13个碱基存在差异,同源性为96%;由碱基序列推定编码的氨基酸残基序列没有发生变异,同源性为100%。结论:成功地将HpHspA基因克隆到了pGEX4T-1原核表达载体,为HspA的表达和研究奠定了实验基础。

Objective： To get heat shock protein A （HspA） subunit gene of Helicobacter pylori, analyze its DNA sequence, and compare its homology. Methods： Chromosome DNAof H. pylori was extracted and the HspA gene was amplified with Polymerase Chain Reaction （PCR）. The obtained DNA fragment was cloned into prokaryotic expression vector pGEX-4T-1 and sequenced. Results： HspA gene was cloned successfully which was demonstrated by enzyme digesting and gene sequencing methods. Comparing with DNA sequences reported in Genbank, nucleotide sequence of the obtained DNA fragment was 96% identical with that of HspA gene with a variation of 13 base pairs. The homology in putative amino acids was 100%. Conclusions： HspA gene has been Cloned into prokaryotic expression vector pGEX 4T-1 successfully, which makes an experimental foundation for its expression and associated research.