[腺病毒载体转染的哺乳动物细胞中乙肝病毒前S2/S基因的表达]
[Expression of HBV preS2/S gene in mammalian cells transferred with adenoviral vector].
作者信息
Lei Chun-liang, Huang Cheng-hui, Yang Zhan, Tang Xiao-ping
机构信息
The Eighth Hospital of Guangzhou, Guangzhou 510060, China.
出版信息
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2005 Mar;19(1):55-7.
OBJECTIVE
To study HBV preS2/S gene expression effects in mammalian cells transferred with recombinant adenoviral vector.
METHODS
The replication-deficient recombinant adenoviral vector (Ad-HBs) carrying HBV preS2/S gene were constructed by homologous recombination in bacteria. The 293 cells, Vero cells, HepG2 cells and mesenchymal stem cells (MSCs) were infected with adenoviruses. The expressions of enhanced green fluorescent protein (EGFP) were observed with fluorescence microscope and the expressions of HBsAg were detected by RT-PCR and ELISA in vitro.
RESULTS
More than 90% of 293 cells, Vero cells, HepG2 cells or MSCs expressed EGFP after transfection at the MOI of 20 and the titers of HBsAg were more than 3.229 (A value) in culture supernatant.
CONCLUSION
The HBV preS2/S gene was not only expressed efficiently in immortalized cells, but also expressed efficiently in stem cells with the recombinant adenoviruses vector.
目的
研究重组腺病毒载体转染哺乳动物细胞后乙肝病毒前S2/S基因的表达效果。
方法
通过细菌中的同源重组构建携带乙肝病毒前S2/S基因的复制缺陷型重组腺病毒载体（Ad-HBs）。用腺病毒感染293细胞、Vero细胞、HepG2细胞和间充质干细胞（MSCs）。用荧光显微镜观察增强型绿色荧光蛋白（EGFP）的表达，并通过RT-PCR和ELISA体外检测HBsAg的表达。
结果
在感染复数（MOI）为20转染后，超过90%的293细胞、Vero细胞、HepG2细胞或MSCs表达EGFP，培养上清中HBsAg滴度超过3.229（A值）。
结论
乙肝病毒前S2/S基因不仅能在永生化细胞中高效表达，而且通过重组腺病毒载体在干细胞中也能高效表达。
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