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含CD基因的人大肠癌特异性逆转录病毒载体的构建
AIM In order to express suicide gene specifically in colorectal carcinoma cells, we constructed the recombinant retroviral vector promoted by transcriptional regulatory sequences of carcinoembryonic antigen (CEA) gene.METHODS A 1.3 kb EcoRⅠ/ BamHⅠ fragment of the CD gene was inserted into pUC118, and then the HindⅢ/ EcoRⅠ CD fragment was isolated. A 420 bp HindⅢ/ BamHⅠ fragment from pCEA 424/ 2CAT was released and subcloned into pUC118 to harvest HindⅢ/ EcoRⅠ CEA promoter. These two fragments were ligated with the linearized retroviral vector GINa using T4 DNA ligase.RESULTS Restriction enzyme SalⅠ was used to digest the recombinants to identify a positive clone A7 and a 1.7 kb band was observed on agarose electrophoresis gel. The recombinant retroviral vector, GICEACDNa was constructed.CONCLUSION Success in construction of GICEACDNa provides a basis for colorectal carcinoma-specific gene therapy.
