Construction of a liver targeting gene transfer vector using hepatitis B virus envelope particles and assessment of its function
- VernacularTitle:作为基因转移载体的乙肝病毒衣壳的制备及其功能的初步鉴定
- Author:
Dejian PAN
;
Zhongquan ZHAO
;
Donglin WANG
;
Zhengtang CHEN
- Publication Type:Journal Article
- Keywords:
Hepatitis B virus, envelope, liver, targeting, gene transfer
- From:
Chinese Journal of Cancer Biotherapy
2006;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
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 ?-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES_2-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 S_1+pre S_2, 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 ?-propiolactone.
