Construction and expression of eukaryotic expression vector for hepatitis B virus X gene containing EGFP

WANG Wenlong; et al

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Construction and expression of eukaryotic expression vector for hepatitis B virus X gene containing EGFP

更新时间：2024-01-17

- Construction and expression of eukaryotic expression vector for hepatitis B virus X gene containing EGFP
- Journal of Chongqing Medical University Issue 5, Pages: 652-654(2006)
- 作者机构：
  
  1. 中南大学湘雅二医院肝病研究中心
  2. 中南大学湘雅二医院肝病研究中心,湖南,长沙,410011
- 作者简介：
- 基金信息：
- DOI：
  CLC： R373
- Published：2006，
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[1]王文龙,贺兴鄂,杨旭,蒋永芳,雷建华,孙会卿.乙肝病毒X基因真核表达载体pEGFP-X的构建及其表达[J].重庆医科大学学报,2006(05):652-654+680.

WANG Wenlong, et al. Construction and expression of eukaryotic expression vector for hepatitis B virus X gene containing EGFP. [J]. Journal of Chongqing Medical University (5):652-654(2006)
[1]王文龙,贺兴鄂,杨旭,蒋永芳,雷建华,孙会卿.乙肝病毒X基因真核表达载体pEGFP-X的构建及其表达[J].重庆医科大学学报,2006(05):652-654+680. DOI：

WANG Wenlong, et al. Construction and expression of eukaryotic expression vector for hepatitis B virus X gene containing EGFP. [J]. Journal of Chongqing Medical University (5):652-654(2006) DOI：

摘要

目的:构建pEGFP-X真核表达载体

观察其在肝癌细胞株中的表达。方法:从质粒pcDNA3.1（+）-X酶切获HBV X基因片段

克隆至pEGFP-N1;用酶切、PCR和测序鉴定pEGFP-X载体;用脂质体法

将重组载体（pEGFP-X）和空载体（pEGFP-N1）瞬时转染肝癌细胞细胞株BEL-7402;分别用RT-PCR、荧光显微镜鉴定HBV X和EGFP基因表达。结果:鉴定证实pEGFP-X载体构建成功;该质粒瞬时转染的BEL-7402有HBV X基因表达

并发绿色荧光。结论:构建的pEGFP-X真核载体能在BEL-7402中表达

绿色荧光蛋白示踪利于表达HBV X基因稳定细胞株的筛选

并为探讨HBV X基因在HCC中的致瘤机理提供实验模型奠定基础。

Abstract

Objective:To construct eukaryotic expression vector for hepatitis B virus X(HBV X) gene with enhanced green fluorescence protein(EGFP)

and confirm its expression in hepatocellular carcinoma(HCC)cell line.Method: HBV X gene was cloned from pcDNA3.1(+)-X by enzyme digestion and inserted to pEGFP-N1.The rector was confirmed by enzyme digestion

PCR assay and sequencing.Then pEGFP-X was transfected to HCC cell line Bel-7402.After transient transfection

expressions of HBV X and EGFP gene were detected by RT-PCR and fluorescence microscope respectively.Results: The results showed that the recombinant plasmid could express HBV X gene efficiently in Bel-7402

which showed green fluorescence.Conclution: The pEGFP-X was constructed and the fused HBV X-EGFP gene was expressed in Bel-7402 successfully.These facilitate the study of the effect of HBV X protein on the development of HCC.

关键词

乙型肝炎病毒X基因加强型绿色荧光蛋白真核表达载体RT-PCRBel-7402

Keywords

Hepatitis B virus X geneEGFPeukaryotic expression vectorRT-PCRBel-7402

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