构建表达Flag-GPI的重组乙型肝炎病毒载体

胡孔颖1; 谷陈建1; 吴敏乐2; 陶帅1; 刘楠楠1; 刘晶1; 谢幼华1

doi:10.3969/j.issn.1673-6184.2019.04.002

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微生物与感染 ›› 2019, Vol. 14 ›› Issue (4) : 199-208. DOI: 10.3969/j.issn.1673-6184.2019.04.002

论著

构建表达Flag-GPI的重组乙型肝炎病毒载体

胡孔颖¹,谷陈建¹,吴敏乐²,陶帅¹,刘楠楠¹,刘晶¹,谢幼华¹

作者信息 +

Construction of Flag-GPI carrying recombinant hepatitis B virus vector

HU Kongying¹, GU Chenjian¹, WU Minle², TAO Shuai¹, LIU Nannan¹, LIU Jing¹, XIE Youhua1

Author information +

文章历史 +

摘要

乙型肝炎病毒(hepatitis B virus，HBV)是小型的嗜肝DNA病毒，能够特异性地感染人肝实质细胞。HBV的易感性可以归于多个方面，包括细胞表面的受体以及细胞内的蛋白或其他因子，目前对HBV易感性的了解还有待深入。本课题利用课题组原有的HBV 5c3c载体和糖基磷脂酰肌醇(glycosylphosphatidylinositol，GPI)的特性，构建了重组HBV载体5c3c-CD59-Flag-GPI和5c3cT-Flag-GPI，转染Huh7细胞后可以将Flag标签锚定在细胞膜上，且可利用Flag抗体通过流式细胞术对其进行分选。在回补HBV包膜蛋白的情况下，5c3cT-Flag-GPI能包装形成完整的重组HBV颗粒。本研究为后续优化重组HBV的包装效率，进而为HBV易感性研究提供工具并奠定基础。

Abstract

Hepatitis B virus (HBV) is a small hepatophilic DNA virus that can specifically infect human hepatic parenchymal cells. The susceptibility of HBV can be attributed to cell surface receptor(s), intracellular proteins and other factors. A simple and clear method to assay the susceptibility of cells for HBV remains a challenge. Here, we constructed the recombinant HBV vectors 5c3c-CD59-Flag-GPI and 5c3cT-Flag-GPI based on the original HBV 5c3c vector and glycosylphosphatidylinositol (GPI) anchor. After transfection into Huh7 cells, Flag tags can be anchored on the cell membrane, and sorted by flow cytometry using Flag antibody. With the replenishment of HBV envelope protein, 5c3cT-Flag-GPI can form complete recombinant HBV particles. This study laid a foundation for optimization of packaging efficiency of recombinant HBV, which would provide a tool for HBV susceptibility research.

导出引用

胡孔颖1，谷陈建1，吴敏乐2，陶帅1，刘楠楠1，刘晶1，谢幼华1. 构建表达Flag-GPI的重组乙型肝炎病毒载体[J]. 微生物与感染. 2019, 14(4): 199-208 https://doi.org/10.3969/j.issn.1673-6184.2019.04.002

HU Kongying1, GU Chenjian1, WU Minle2, TAO Shuai1, LIU Nannan1, LIU Jing1, XIE Youhua1. Construction of Flag-GPI carrying recombinant hepatitis B virus vector[J]. Journal of Microbes and Infections. 2019, 14(4): 199-208 https://doi.org/10.3969/j.issn.1673-6184.2019.04.002