Construction and expression of the eukaryotic expression plasmid  encoding mycobacterium tuberculosis MPT64

LUO Xudong; et al

doi:10.13406/j.cnki.cyxb.2003.02.001

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Construction and expression of the eukaryotic expression plasmid encoding mycobacterium tuberculosis MPT64

更新时间：2024-01-17

- Construction and expression of the eukaryotic expression plasmid encoding mycobacterium tuberculosis MPT64
- Journal of Chongqing Medical University Issue 2, Pages: 125-126(2003)
- 作者机构：
  
  1. 重庆医科大学微生物教研室
  2. 重庆医科大学微生物教研室,重庆,400016
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2003.02.001
  CLC： R346
- Published：2003，
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[1]骆旭东,陈全,蒋英,江山,朱道银.结核分枝杆菌MPT64真核表达质粒的构建及其表达[J].重庆医科大学学报,2003(02):125-126+168.

LUO Xudong, et al. Construction and expression of the eukaryotic expression plasmid encoding mycobacterium tuberculosis MPT64. [J]. Journal of Chongqing Medical University (2):125-126(2003)
[1]骆旭东,陈全,蒋英,江山,朱道银.结核分枝杆菌MPT64真核表达质粒的构建及其表达[J].重庆医科大学学报,2003(02):125-126+168. DOI： 10.13406/j.cnki.cyxb.2003.02.001.

LUO Xudong, et al. Construction and expression of the eukaryotic expression plasmid encoding mycobacterium tuberculosis MPT64. [J]. Journal of Chongqing Medical University (2):125-126(2003) DOI： 10.13406/j.cnki.cyxb.2003.02.001.

摘要

目的 :构建结核分枝杆菌MPT6 4真核表达质粒

并在真核细胞中表达。方法 :从H37Rv基因组中扩增出MPT6 4基因

经限制性内切酶消化后

定向插入 pcDNA3.1（+）中

用脂质体法将 pcDNA -MPT6 4转染COS - 7细胞

采用RT -PCR、ELISA和斑点印迹法检测其表达。结果 :扩增出的MPT6 4基因正确插入pcDNA3.1中

DNA序列测定无突变产生。重组质粒在COS - 7细胞中表达了MPT6 4。结论 :成功地构建了 pcDNA -MPT6 4质粒

其真核表达的蛋白具有良好的抗原性

为进一步研究MPT6 4在抗结核菌感染中的预防作用奠定了基础。

Abstract

Objective:To construct the recombinant eukaryotic plasmid encoding Mycobacterium tuberculosis MPT64 and detect its expression in COS-7 cell lines. Methods: The gene encoding MPT64 was amplified by polymerase chain reaction (PCR) from H37Rv genome

and the fragment was inserted into pcDNA3.1(+) after restriction endonuclease digestion .The recombinant plasmid pcDNA MPT64 was transfected into COS-7 cell lines with liposome

and expressed MPT64 in COS-7 cell was detected by RT-PCR

ELISA and dot blotting. Results: The sequence of MPT64 gene in recombinant plasmid was the same as that of reported

and recombinant MPT64 protein was expressed in COS-7 cell lines.Conclusion: The recombinant plasmid pcDNA MPT64 was constructed successfully

and MPT64 protein expressed in COS-7 cell has antigenicity.The results established the basis for further studies on MPT64 function.

关键词

结核分枝杆菌MPT64真核表达质粒

Keywords

mycobacterium tuberculosisMPT64Eukaryotic expression plasmid

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Related Institution

Department of Anesthesiology， Chongqing University Cancer Hospital

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Department of Clinical Immunology,Chongqing Medical University

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