Construction of Apoptin Fusion Gene and Reporter Gene pECFP-C1 and the Establishment of Their Stable Expression Cell Strains

The apoptin fusion gene was cloned and the clone vector pMD-18T-AFG was constructed. And then the apoptin fusion gene was subcloned into the multiclone sites of reporter gene pECFP-C1 between EcoRⅠand SacⅡ to construct the eukaryotic expression vector of apoptin fusion gene with reporter gene. The eukaryotic expression vector of apoptin fusion gene with reporter gene was transfected into Chinese hamster ovary cells (CHO) by lipidosome transfection. The results showed that CHO appeared bright fluorescence 48 h after transfection, indicating that the eukaryotic expression vector of apoptin fusion gene with reporter gene has been constructed and expressed in CHO cells. RT-PCR showed that the apoptin fusion gene was expressed in CHO cells. The stable expression cell line was obtained by G418 screening after 3 weeks. The results were significant for further study on the localization of apoptin fusion protein in tumor tissues and the effects of apoptin fusion protein on apoptosis of cancer cells.