| EGFP-HBVP22e融合蛋白在HepG2中的表达 |
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| 引用本文： | 刁志宏,张明霞,朱幼芙,何海棠,王战会,侯金林. EGFP-HBVP22e融合蛋白在HepG2中的表达[J]. 解放军医学杂志, 2007, 32(1): 61-63 |
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| 作者姓名： | 刁志宏 张明霞 朱幼芙 何海棠 王战会 侯金林 |
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| 作者单位： | 510515,广州,南方医科大学南方医院感染内科;510515,广州,南方医科大学南方医院感染内科;510515,广州,南方医科大学南方医院感染内科;510515,广州,南方医科大学南方医院感染内科;510515,广州,南方医科大学南方医院感染内科;510515,广州,南方医科大学南方医院感染内科 |
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| 摘 要： | 目的 利用pEGFP-C2真核表达载体,建立稳定表达乙型肝炎病毒(HBV)P22e的HepG2细胞系.方法 以含1.2拷贝HBV基因(adr亚型)的p3.8Ⅱ质粒为模板,PCR获得HBVP22e cDNA片段,分离插入通用T载体pMD18-T中鉴定,然后装入真核表达型载体pEGFP-C2中,HBVP22e与EGFP基因融合,脂质体转染HepG2细胞株,荧光显微镜下观察EGFP-HBVP22e融合蛋白的胞内表达,Western blot检测阳性克隆细胞EGFP-HBVP22e的表达.结果 成功构建了真核表达载体pEGFP-C2HBVP22e,并转染HepG2细胞,经多次传代培养鉴定,获得稳定表达EGFP-HBVP22e融合蛋白的HepG2细胞系.结论 该细胞系的建立为进一步研究HBVP22e的生物学功能与乙型肝炎发病机制的关系奠定了基础.
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| 关 键 词： | 肝炎病毒 乙型 基因重组 真核表达 |
| 收稿时间： | 2006-06-19 |
| 修稿时间： | 2006-11-12 |
Expression of fusion protein EGFP-HBVP22e in HepG2 cells |
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| Diao Zhihong, Zhang Mingxia, Zhu Youfu, et al. Expression of fusion protein EGFP-HBVP22e in HepG2 cells[J]. Medical Journal of Chinese People's Liberation Army, 2007, 32(1): 61-63 |
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| Authors: | Diao Zhihong Zhang Mingxia Zhu Youfu et al |
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| Affiliation: | Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China |
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| Abstract: | Objective With eukaryote expression vector pEGFP-C2, to establish the HepG2 cell line which may stably express the P22e of hepatitis B virus (HBV), and may be used to study the existed relationship between HBVP22e and hepatitis B. Methods HBVP22e cDNA obtained by PCR from HBV adr subtype 1.2 copies genome plasmid p3.8II was inserted into universal vector pMD18-T for identification. The plasmids were transfected into HepG2 cells via liposome, while EGFP-HBVP22e was analyzed by Western blot, and observed with fluorescence microscope in HepG2 cells. Results Expression vectors of recombinant pEGFP-C2HBV P22e were constructed and expressed steadily in HepG2 cells. Conclusion The results can be used to explore biological activity of HBVP22e in hepatitis B. |
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| Keywords: | hepatitis B virus gene recombinant eukaryote expression |
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