人幽门螺杆菌热休克蛋白A编码基因的克隆及序列分析

目的：获取含人幽门螺杆菌(Hp)热休克蛋白A编码基因并构建其重组载体，进行核苷酸序列分析，为在E．coli BL21中表达，疫苗的开发奠定基础。方法：利用分子克隆技术从Hp DNA染色体中，扩增热休克蛋白A编码基因片段。将目的基因与pET32a( )同时经kpnI、BamHI双酶切、纯化、连接后。转化含有目的基因的重组载体，进行序列分析。结果：经酶切、测序分析表明，插入的基因片段为Hp热休克蛋白A编码基因，与GenBank的报道相比较，有1．4％的bp发生变异，1．6％的氨基酸残基改变。其同源性高达98％。结论：成功地克隆了Hp热休克蛋白A编码基因，为Hp蛋白质疫苗的研制和快速诊断试剂盒的开发奠定了良好的基础。

Cloning and sequence analysis of human heat shock prote in encoding gene of Helicobacter pylori

Objective:To obtain DNA of human Helicobacter pylori heat shock protein A, and construct a recombinant vector containing gene encoding HspA for nucleotide sequence analysis.Methods:The target gene was amplified from Hp chromosome by PCR, and then digested by restricted endonuclease enzyme of kpn I , BamH I simultanously, and inserted into the prokaryotic expression vector pET32a(+) digested by corresponding restricted endonuclease enzyme. The recombinant vector was selected and transformed for nucleotide sequence analysis.Results:Enzyme digestion analysis and sequencing showed that the target gene had been inserted into recombinant vector, but as compared with gene reported by GenBank. 1.4 % of the gene mutation and 1.6% of amino acid residues change in Hp happened respectively. The DNA sequence analysis showed the sequence of HspA DNA was almost the same as that published by GenBank.Conclusion:The gene coding for Hp HspA is cloned successfully.The results obtained lay the foundation for research on development of Hp protein vaccine and a quickly diagnostic kit applying to detection of Hp infection.