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

JIANG Zheng; et al

doi:10.13406/j.cnki.cyxb.2003.04.003

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

更新时间：2024-01-17

- Cloning and sequence analysis of human heat shock prote in encoding gene of Helicobacter pylori
- Journal of Chongqing Medical University Issue 4, Pages: 414-416(2003)
- 作者机构：
  
  1. 重庆医科大学临床学院消化内科
  2. 重庆医科大学病毒性肝炎研究所
  3. 重庆医科大学临床学院消化内科,重庆,400016
  4. ,重庆,400010
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2003.04.003
  CLC： R346
- Published：2003，
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[1]姜政,黄爱龙,蒲丹,陶小红,王丕龙.人幽门螺杆菌热休克蛋白A编码基因的克隆及序列分析[J].重庆医科大学学报,2003(04):414-416+420.

JIANG Zheng, et al. Cloning and sequence analysis of human heat shock prote in encoding gene of Helicobacter pylori. [J]. Journal of Chongqing Medical University (4):414-416(2003)
[1]姜政,黄爱龙,蒲丹,陶小红,王丕龙.人幽门螺杆菌热休克蛋白A编码基因的克隆及序列分析[J].重庆医科大学学报,2003(04):414-416+420. DOI： 10.13406/j.cnki.cyxb.2003.04.003.

JIANG Zheng, et al. Cloning and sequence analysis of human heat shock prote in encoding gene of Helicobacter pylori. [J]. Journal of Chongqing Medical University (4):414-416(2003) DOI： 10.13406/j.cnki.cyxb.2003.04.003.

摘要

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 .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。

Abstract

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.

关键词

幽门螺杆菌热休克蛋白A编码基因重组载体疫苗

Keywords

Helicobacter pyloriHeat shock protein AEncoding geneRecombinant vectorvaccine

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