Construction of retroviral vector carrying HSV-tk gene under control of human AFP enhancer core sequence and human pgk promotor.

ABSTRACT

AIM: To construct retroviral vector bringing HSV-tk gene under control of human AFP enhancer core sequence and human pgk promoter. METHODS: Internal SV40 promoter was deleted by SalI from retroviral vector pMNSM to construct pMNM. HSV-tk gene driven by pgk promoter was released by BamH I from an eukaryotic expression vector pBPGK-tk, and inserted into polylinker site of pMNM to construct pMNP-tk retroviral vector. Human α-fetoprotein gene enhancer core sequence was released by EcoR I from pGEM. 7Z-AFPe plasmid was inserted into the immediate upstream of pgk promoter of pMNP-tk vector. Construction of hepatoma specific retroviral vector pMNAP-tk was completed. RESULTS: The structure of pMNP-tk and pMNAP-tk vector was confirmed by restriction analysis. CONCLUSION: The vector is of great significance for hepatoma specific prodrug transformation gene therapy.