Construction of plasmid coexpressing hepatitis B surface antigen and granulocyte macrophage-colony stimulating factor

QING Yuling; et al

doi:10.13406/j.cnki.cyxb.2003.03.002

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Construction of plasmid coexpressing hepatitis B surface antigen and granulocyte macrophage-colony stimulating factor

更新时间：2024-01-17

- Construction of plasmid coexpressing hepatitis B surface antigen and granulocyte macrophage-colony stimulating factor
- Journal of Chongqing Medical University Issue 3, Pages: 261-263(2003)
- 作者机构：
  
  1. 重庆医科大学病毒性肝炎研究所
  2. 重庆医科大学病毒性肝炎研究所,重庆,400010
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2003.03.002
  CLC： R346
- Published：2003，
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[1]卿玉玲,赵嘉将,任红.HBsAg和GM-CSF融合表达质粒的构建[J].重庆医科大学学报,2003(03):261-263+266.

QING Yuling, et al. Construction of plasmid coexpressing hepatitis B surface antigen and granulocyte macrophage-colony stimulating factor. [J]. Journal of Chongqing Medical University (3):261-263(2003)
[1]卿玉玲,赵嘉将,任红.HBsAg和GM-CSF融合表达质粒的构建[J].重庆医科大学学报,2003(03):261-263+266. DOI： 10.13406/j.cnki.cyxb.2003.03.002.

QING Yuling, et al. Construction of plasmid coexpressing hepatitis B surface antigen and granulocyte macrophage-colony stimulating factor. [J]. Journal of Chongqing Medical University (3):261-263(2003) DOI： 10.13406/j.cnki.cyxb.2003.03.002.

摘要

目的 :构建乙肝HBsAg和GM -CSF的融合表达质粒

借助GM -CSF的免疫佐剂作用

提高乙型肝炎DNA疫苗的免疫效果。方法 :用PCR方法分别从 pEcob6和 pCD -hGM -CSF中扩增出基因S和GM -CSF

克隆到真核表达质粒pcD NA3.1（+）中

构建pcDNA3.1-S及融合表达质粒 pcDNA3.1-GM -CSF -S

然后以重组质粒转染真核细胞

研究重组质粒体外表达。结果 :经酶切鉴定及DNA序列证实重组质粒构建正确

细胞转染试验表明重组质粒能在COS - 7及HepG - 2内表达。结论 :乙肝HBsAg和GM -CSF的融合表达质粒构建成功

重组质粒能在真核细胞内表达。

Abstract

Objective:To construct plasmid coexpressing GM-CSF and HBsAg

enhance hepatitis B DNA vaccines antivirus effect.Methods:(1)The HBsAg-encoding fragment and GM-CSF-encoding fragment were created by PCR amplification from pEcob6 and pCD-hGM-CSF respectively

then were cloned into plasmid pcDNA3.1(+)

pcDNA3.1-S and pcDNA3.1-GM-CSF(without stop codon)were constructed.(2)HBsAg DNA fragment (with stop codon) was amplified by PCR

then was subcloned into plasmid pcDNA3.1-GM-CSF.Results:Recombinant pcDNA3.1-S and pcDNA3.1-GM-CSF-S were confirmed by using restriction enzymes and DNA sequencing.HBsAg was detected in the lysates and supernatants from cells transfected with pcDNA3.1-S and in the lysates from cells transfected with pcDNA3.1-GM-CSF-S.Conclusion:Two eukaryotic expression plasmids were constructed successfully and could express HBsAg.

关键词

乙型肝炎粒细胞、巨噬细胞集落刺激因子质粒构建

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