目的： 构建具有肝脏靶向性的乙型肝炎病毒衣壳基因转移载体。方法: 采用PEG 8 000病毒浓缩法提取HepG 2.2.15细胞上清液中的乙型肝炎病毒，用β 丙内酯法制备乙型肝炎病毒衣壳，用它包裹5.3kb的绿色荧光蛋白质粒以测试其装载能力，并用ELISA法、PCR法、SDS 聚丙烯酰胺凝胶电泳、透射电镜等对它进行定量、定性研究，最后将其转染HepG 2细胞，通过荧光显微镜观察绿色荧光蛋白表达情况，通过流式细胞仪检测细胞发光率。结果: 制备的乙型肝炎病毒衣壳载体保留病毒衣壳的表面蛋白HBsAg+pre S1+pre S2，无病毒DNA；该载体对绿色荧光蛋白质粒较高装载容量，装载绿色荧光蛋白质粒后在肝癌细胞中有很高的转移效率，并能实现高表达。结论: 采用PEG 8 000病毒浓缩法、β 丙内酯法可从HepG2215细胞上清液中制备乙型肝炎病毒衣壳基因转移载体，该载体具有良好的生物学功能。

Objective: To construct a liver targeting gene transfer vector using hepatitis B virus envelope particles. Methods: Hepatitis B viruses were obtained from the supernatant of HepG 2.2.15 cells by a PEG8000 system and were inactivated by β-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES2-EGFP to assess their packing ability. Subsequently, the products were studied with ELISA, PCR, SDS-PAGE, and electron microscopy. Finally, the product was used to transfect HepG2 cells and the green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results: The acquired hepatitis B virus envelope retained the surface protein HBsAg+pre S1+pre S2, but with no virus DNA. The prepared envelope had high packing ability for GFP and the packed GFP had a high transfection rate in HepG2 cell. Conclusion: Hepatitis B virus envelope has been successfully obtained from the supernatant of HepG 2215 cells with a PEG8000 system and β-propiolactone.

国家自然科学基金资助项目（No.30100189）