HCV C基因腺病毒表达载体骨架质粒pAd.HCV-C的构建、鉴定及表达

Construction, identification and expression of framework plasmid pAd. HCV-C of adenovirus expression vector of HCV C

AIM To construct a recombinant framework plasmid of adenovirus expression vector which can express HCV core gene. This should be useful to pack adenovirus expression vector which can express HCV core gene effectively. METHODS The HCV core gene derived from the plasmid pBRTM/HCV 1-3011 by using polymerase chain reaction (PCR) was inserted into the backward position of cytomegalovirus (CMV) immediate early promotor element of framework plasmid (pAd. CMV-Link. 1) of adenovirus expression vector, then the recombinant plasmid pAd. HCV-C was obtained. The pad. HCV-C was identified by endonuclease, PCR and sequencing. HCV core gene was expressed transiently with Lipofectamine coated in human hepatoblastoma 7721 cells which examined by indirect immunofluorescence. RESULTS Insert DNA of pAd. HCV-C was confirmed HCV core gene by endonuclease, PCR and sequencing, pAd. HCV-C can express HCV core gene transiently in 7721 cells which confirmed by indirect immunofluorescence. CONCLUSION Recombinant framework plasmid of adenovirus expression vector pAd. HCV-C can express HCV core gene transiently in 7721 cells. This should be useful to pack adenovirus expression vector which can express HCV core gene.