Construction and identification of recombinant adenovirus containing double suicide fusion gene

Objective:To construct a recombinant adenovirus containing cytosine deaminase( CD ) gene and thymidine kinase( TK ) fusion gene for the gene therapy research of malignant tumors. Methods: A recombinant cosmid containing CD and TK fusion gene was constructed, and then mixed with DNA TPC and co transfected to the 293 cells by calcium phosphate coprecipitation. Results: The results of restriction and PCR showed the insertion was right and the recombinant adenovirus generated contained the CD and TK fusion gene without replication competent adenovirus. Conclusion: The recombinant adenovirus generated is E1 and E3 deleted and contains double suicide gene needed, which can be further studied for gene therapy.