Study on prokaryotic expression of hepatitis C virus P7 protein

CHEN Ke; et al

doi:10.13406/j.cnki.cyxb.2011.03.001

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Study on prokaryotic expression of hepatitis C virus P7 protein

更新时间：2024-01-17

- Study on prokaryotic expression of hepatitis C virus P7 protein
- Journal of Chongqing Medical University Vol. 36, Issue 3, Pages: 262-265(2011)
- 作者机构：
  
  重庆医科大学感染性疾病分子生物学教育部重点实验室
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2011.03.001
  CLC： R373
- Published：2011，
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[1]陈可,胡接力,胡源,张文露,王增产,汤华,黄爱龙,蔡雪飞.丙型肝炎病毒P7蛋白体外原核表达状况的研究[J].重庆医科大学学报,2011,36(03):262-265.

CHEN Ke, et al. Study on prokaryotic expression of hepatitis C virus P7 protein. [J]. Journal of Chongqing Medical University 36(3):262-265(2011)
[1]陈可,胡接力,胡源,张文露,王增产,汤华,黄爱龙,蔡雪飞.丙型肝炎病毒P7蛋白体外原核表达状况的研究[J].重庆医科大学学报,2011,36(03):262-265. DOI： 10.13406/j.cnki.cyxb.2011.03.001.

CHEN Ke, et al. Study on prokaryotic expression of hepatitis C virus P7 protein. [J]. Journal of Chongqing Medical University 36(3):262-265(2011) DOI： 10.13406/j.cnki.cyxb.2011.03.001.

摘要

目的:扩增丙型肝炎病毒（Hepatitis Cvirus

HCV）P7蛋白基因

克隆到多个原核表达系统

观察P7蛋白在体外表达的情况

获取具有生物学活性的HCVP7重组蛋白。方法:设计扩增全长HCVP7基因片段的特异引物

以H/FL质粒DNA（含HCV1acDNA全长序列）为模板

通过PCR扩增全长HCV P7基因

定向克隆到pQE30、pGEX 4T-2、pET32a（+）3种不同类型的原核表达载体中。将重组后包含HCVP7基因的原核表达载体转化表达型大肠杆菌BL21（DE3）pLysS或SG13009（PREP4）

并做异丙基硫化-β-D-半乳糖苷（Isopropyl-beta-D-thiogalactoside

IPTG）诱导表达

通过SDS-PAGE和Western blot鉴定HCV P7蛋白原核表达的情况。结论:获得可溶性的重组蛋白GST-HCVP7和Trx-HCVP7

为进一步研究P7蛋白的生物学功能奠定基础。

Abstract

Objective: To amplify HCV P7 gene and clone it into several prokaryotic expression systems

then investigate the effects of these working systems and harvest HCV P7 protein with biologic activity.Methods: Design Special primers were designed for the amplification of HCV P7 complete DNA sequence and amplify P7 DNA fragments were amplified with H/FL plasmid as template by PCR.The obtained P7 DNA fragments were directly cloned into pQE30

pGEX 4T-2 and pET32a(+) prokaryotic expression vectors and transform these recombinant plasmids were transformed into E coli.strain BL21(DE3) pLysS or SG13009(PREP4)

and IPTG was added to induce recombinant E coli.expression.The expression products were identified with SDS-PAGE and Western blot.Results: There was no track of non-fusion HCV P7 protein expression from recombinant E coli

including pQE30/HCV P7 or pET32a(+)-Nde/HCV P7 plasmid.But the study succeeded in the expression and purification of recombinant HCV P7 proteins

Both are all soluble and biologic activity

and are expected to use extensively in anti-HCV virus research.

关键词

丙型肝炎病毒P7蛋白原核表达

Keywords

HCVP7 proteinprokaryotic expression

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