摘要
目的 获取含人幽门螺杆菌 (Hp)尿素酶B编码基因并构建其原核表达的重组载体 ,进行核甘酸序列分析 ,为在E .coliBL2 1中表达 ,疫苗的开发奠定基础。方法 利用分子克隆技术从HpDNA染色体中 ,扩增尿素酶B编码基因片段。将目的基因与pET32a(+)同时经SacI、Xhol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尿素酶B编码基因 ,与GenBank公布的序列相比较 ,有 1.8%的bp发生变异 ,0 .35 %的氨基酸残基改变 ,但同源性高达 98%。结论 成功地克隆了Hp尿素酶B编码基因 。
Objective To obtain DNA of human Helicobacter pylori urease B, and construct a recombinant vector containing gene encoding urease B for nucleotide sequence analysis. Methods The target gene was amplified from Hp chromosome by PCR. And then digested by restricted endonuclease enzyme of SacI, Xhol simultanously, and inserted into the prokaryotic expression vector pET32a(+) digested by corresponding restricted endonuclease enzyme. The recombinant vector was used to select and transform 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, 1.8 % of the gene mutation and 0.35 % of amino acid residues change in Hp happened respectively. The DNA sequence analysis showed the sequence of urease B DNA was almost same as that of the published by GenBank. Conclusion The gene coding for Hp urease B 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.
出处
《重庆医学》
CAS
CSCD
2003年第7期865-867,共3页
Chongqing medicine
关键词
幽门螺杆菌
尿素酶B
原核表达栽体
编码基因
helicobacter pylori
urease B
the prokaryotic expression vector
encoding gene
