乙型肝炎病毒S基因重组逆转录病毒载体的构建及其在真核细胞中表达  

Construction of the recombinant retrovirus vector of HBV-S gene and it's expression in eukaryotic cells

作　　者：周智[1] 张定凤[2] 任红[2] 

机构地区：[1]中山医科大学附属第三医院传染科肝病室,广州510630 [2]重庆医科大学病毒性肝炎研究所,400010

出　　处：《中华肝脏病杂志》2000年第5期296-298,共3页Chinese Journal of Hepatology

基　　金：国家自然科学基金!367634;卫生部课题基金

摘　　要：目的探索逆转录病毒载体在基因治疗中的应用。方法用DNA重组技术构建HBV－S基因重组逆转录病毒载体，电穿孔转染PA317后，筛选高表达克隆，用假病毒颗粒感染HepG_2、P815和EL4细胞，分别用RT－PCR及ELISA法检测目的基因表达。结果HBsAg在上述细胞中获得不同程度的表达，细胞上清液（48h）中HBsAg含量（吸光度值）分别为0.92、0.09和0.47结论逆转录病毒用载体，其目的基因在细胞内稳定表达，是一种高效的基因转移系统，适用于基因治疗实验。Objective To investigate the effectiveness of recombinant retrovirus vector in gene therapy. MethodsThe retroviral vector PLXSN-S was constructed and transferred into PA317 by means of electroporation, then HepG_2, P815,and EL4 cells were infected with the pseuovirus produced from PA317, which highly expressed HBsAg. HBsAg expressionwas tested by RT-PCR and ELISA. Results HBsAg was expressed variously in the eukaryotic cells mentioned above.HBsAg (A value) of the cell supernatants (48 h) were 0.92, 0.09. 0.47, respectively. Conclusion The vector used in thisstudy is an effective one to carry genes of interest to target cells and it may be useful in the test for gene therapy.[

关 键 词：基因重组 真核表达 乙型肝炎 基因治疗 HBV-C 

分 类 号：R512.6[医药卫生—内科学]