The cloning and recombinant vector constructing of lower-weight outer membrane protein encoding gene of Helicobacter pylori

JIANG Zheng; et al

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The cloning and recombinant vector constructing of lower-weight outer membrane protein encoding gene of Helicobacter pylori

更新时间：2024-01-17

- The cloning and recombinant vector constructing of lower-weight outer membrane protein encoding gene of Helicobacter pylori
- Journal of Chongqing Medical University Issue 3, Pages: 243-246(2002)
- 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：
  CLC： R346
- Published：2002，
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[1]姜政,蒲丹,黄爱龙,王丕龙,陶小红.幽门螺杆菌低分子质量外膜蛋白编码基因的克隆与重组载体构建[J].重庆医科大学学报,2002(03):243-246.

JIANG Zheng, et al. The cloning and recombinant vector constructing of lower-weight outer membrane protein encoding gene of Helicobacter pylori. [J]. Journal of Chongqing Medical University (3):243-246(2002)
[1]姜政,蒲丹,黄爱龙,王丕龙,陶小红.幽门螺杆菌低分子质量外膜蛋白编码基因的克隆与重组载体构建[J].重庆医科大学学报,2002(03):243-246. DOI：

JIANG Zheng, et al. The cloning and recombinant vector constructing of lower-weight outer membrane protein encoding gene of Helicobacter pylori. [J]. Journal of Chongqing Medical University (3):243-246(2002) DOI：

摘要

目的:构建幽门螺杆菌外膜蛋白编码基因重组载体

为的疫苗开发、快速诊断试剂盒的研究奠定基础。18kDaHp方法:用从染色体中扩增外膜蛋白的编码基因片段

将目的基因、质粒同时经、Ⅲ双酶PCRHpDNA18kDapQE30BamHIHind切、纯化、连接、转染以及筛选含插入片段的重组载体。结果:经酶切、测序分析插入的基因片段为表达外膜蛋白Hp18kDa编码基因

有碱基发生变异

以及氨基酸残基改变。与文献相比有高度的同源性。2%1.68%

结论:外膜蛋白编码基因克Hp隆、重组载体的构建为该基因编码的蛋白质有效表达提供了条件

同时也为疫苗开发和快速诊断试剂盒的制备打下了基础。

Abstract

Objective: To construct a recombinant vector which could express 18kDa outer membrane protein (OMP) from Helicobacter pylori (Hp)

and explore the possibility to obtain the vaccine conferring protection from Hp infection and a diagnostic reagent kit quickly detecting Hp infection. Methods: The gene encoding the structural 18kDa outer membrane protein of Hp was amplified from Hp chromosomal DNA by PCR

and inserted in the prokaryotic expression vector pQE30. The recombinant vector was transformed into the Top10 E. coli strain

а()selected and identified in order to be transformed into DH5PREP4strain and expressed recombinant protein. Results: The gene of 18kDa OMP was amplified to be 528 base pairs

compared with Toom et al reports: 2% of the gene was mutated

1.68% of amino acid residues was changed

but the homogenicity was about 98% between them. Conclusion: This result suggested that recombinant vector expressed 18kDa OMP may be a potential source for the vaccine and diagnostic reagent kit of Hp.

关键词

幽门螺杆菌外膜蛋白基因原核表达

Keywords

Helicobacter pyloriOuter membrane protein geneprokaryotic expression

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