用结核分枝杆菌HSP70启动子构建分枝杆菌穿梭表达质粒的研究

陈全; 朱道银; 骆旭东; 蒋英; 江山

doi:10.13406/j.cnki.cyxb.2003.06.005

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用结核分枝杆菌HSP70启动子构建分枝杆菌穿梭表达质粒的研究

更新时间：2024-01-17

- 用结核分枝杆菌HSP70启动子构建分枝杆菌穿梭表达质粒的研究
- Construction of the Mycobacterium shuttle expression plasmid with Mycobacterium tuberculosis heat shock protein 70 promoter
- 重庆医科大学学报 2003年第6期页码：697-700
- 作者机构：
  
  1. 重庆医科大学基础医学院微生物学教研室
  2. 重庆医科大学基础医学院微生物学教研室,重庆,400016
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2003.06.005
  中图分类号： R346
- 纸质出版日期：2003，
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[1]陈全,朱道银,骆旭东,蒋英,江山.用结核分枝杆菌HSP70启动子构建分枝杆菌穿梭表达质粒的研究[J].重庆医科大学学报,2003(06):697-700.

CHEN Quan, et al. Construction of the Mycobacterium shuttle expression plasmid with Mycobacterium tuberculosis heat shock protein 70 promoter[J]. Journal of Chongqing Medical University, 2003,(6):697-700.
[1]陈全,朱道银,骆旭东,蒋英,江山.用结核分枝杆菌HSP70启动子构建分枝杆菌穿梭表达质粒的研究[J].重庆医科大学学报,2003(06):697-700. DOI： 10.13406/j.cnki.cyxb.2003.06.005.

CHEN Quan, et al. Construction of the Mycobacterium shuttle expression plasmid with Mycobacterium tuberculosis heat shock protein 70 promoter[J]. Journal of Chongqing Medical University, 2003,(6):697-700. DOI： 10.13406/j.cnki.cyxb.2003.06.005.

摘要

目的 :用结核分枝杆菌HSP70 （Heatshockprotein70

HSP70 ）启动子改建分枝杆菌穿梭质粒 pJEM11为分枝杆菌穿梭表达质粒。方法 :利用聚合酶链反应（Polymerasechainreaction

PCR）扩增H37Rv基因组 4 196 0 7～ 4 19835位的HSP70启动子及调控序列

末端引入一个多克隆位点（Multipleclonesites

MCS）

再定向克隆入 pJEM 11的ApaI位点与XbaI位点之间

构建pJCH0 2载体

并对载体进行酶切鉴定及序列测定。将lhp -esat6融合基因克隆入 pJCH0 2载体

评价其在BCG中的表达情况。结果 :HSP70启动子、调控序列及引入的多克隆位点完全正确

lhp -esat6基因在BCG中成功表达。结论 :成功改建穿梭质粒 pJEM11为穿梭表达质粒 pJCH0 2。本研究为构建BCG多价疫苗奠定了基础.

Abstract

Objective:To reconstruct Mycobacterium shuttle expression plasmid with Mycobacterium tuberculosis HSP70 promoter based on shuttle plasmid pJEM11.Methods:The cDNA fragment of 229bp promoter and its correlative sequence was amplified by PCR.Multiple clone sites was added to the end of amplified fragment

and then it was cloned into the pJEM11 to construct the plasmid pJCH02.The pJCH02 was identified by digestion with restriction endonuclease and sequence analysis.The lhp-esat6 fusion gene was cloned into the pJCH02

then its expression in BCG was confirmed by SDS-PAGE.Results:The sequence of HSP70 promoter and its correlative sequence was in accordance with the predicted sequence.The lhp-esat6 fusion gene was expressed in BCG.Conclusion:The shuttle expression plasmid pJCH02 is constructed successfully.The study provides the basis for BCG polyvaccine.

关键词

结核分枝杆菌热休克蛋白启动子基因表达

Keywords

Mycobacterium tuberculosis(MTB)heat shock proteinpromotergene expression

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