腺病毒载体介导HBsAg基因在哺乳动物细胞中的表达

Expression of HBV preS_2/S gene in mammalian cells transferred with adenoviral vector

目的 探讨腺病毒载体介导乙型肝炎病毒 (HBV)前S2 S(preS2/S)基因在几种哺乳动物细胞中的表达。方法 制备携带HBVpreS2/S基因的非复制型重组腺病毒Ad-HBs ,体外转染人胚肾细胞 (2 93)、绿猴肾细胞 (Vero)、肝癌细胞系 (HepG2 )和间质干细胞 (MSCs) ,荧光显微镜观察EGFP的表达 ,同时采用RT-PCR检测目的基因的转录 ,ELISA法检测目的基因的表达。结果 MOI为 2 0的重组腺病毒Ad-HBs转染 2 93、Vero、HepG2 细胞和MSCs ,4 8h后 90 %以上的细胞表达EGFP ,同时细胞表达高滴度的HBsAg(A值 >3 2 2 9)。结论 腺病毒载体不仅能够介导HBVpreS2 S基因于连续细胞系细胞中高效表达 ,也能介导HBVpreS2

Objective To study HBV preS_2/S gene expression effects in mammalian cells transferred with recombinant adenoviral vector. Methods The replication-deficient recombinant adenoviral vector (Ad-HBs) carrying HBV preS_2/S gene were constructed by homologous recombination in bacteria. The 293 cells, Vero cells, HepG_2 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, HepG_2 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 preS_2/S gene was not only expressed efficiently in immortalized cells, but also expressed efficiently in stem cells with the recombinant adenoviruses vector.