Adeno-associated virus mediated expression of human erythropoietin in vitro  

腺相关病毒介导的人红细胞生成素体外表达(英文)

作　　者：刁勇[1] 许瑞安[2] 王广基[1] 赵小晨 

机构地区：[1]中国药科大学新中新药研究中心,南京210009 [2]香港大学分子生物学研究所

出　　处：《Acta Pharmacologica Sinica》2002年第1期55-58,共4页中国药理学报（英文版）

摘　　要：To investigate the expression of human crythro-poietin (hEpo) in 293 cell line using adeno-associated virus ( AAV) vector, and provide information for its potential application in human gene therapy. METHODS: The human Epo cDNA was inserted into the AAV cassettes, and recombinant adeno-associated virus ( rAAV-EPO) vectors were generated by cotransfec-tion methods without helper virus. The expression of hEpo was detected by RT-PCR and ELISA methods. RESULTS: A recombinant adeno-associated virus expressing hEpo was constructed and generated successfully. Positive signal of mRNA was detected from 293 cells transduced with rAAV-EPO, and the hEpo level in supernatant reached .500 U/L. CONCLUSION: rAAV-EPO expressed biologically active hEpo effectively, suggesting that it is a suitable gene expression tool for further study in vivo experiments.目的:研究腺相关病毒载体介导的人红细胞生成素在293细胞的体外表达Ⅰ方法:将人红细胞生成素cDNA克隆至AAV表达载体,采用无辅助病毒共转染法生产重组AAV-EPO病毒,利用RT-PCR和ELISA法检测hEpo的体外表达.结果:成功构建和生产了表达hEpo的重组AAV-EPO病毒.重组AAV-EPO病毒感染293细胞后,RT-PCR结果显示有hEpo基因表达,细胞上清中hEpo的表达水平达500U/L.结论:所制备的重组AAV-EPO病毒可以有效表达hEpo,可以进一步用于动物体内实验.

关 键 词：DEPENDOVIRUS ERYTHROPOIETIN gene expression gene therapy 

分 类 号：R346[医药卫生—基础医学]