Construction of replication-competent adenovirus shuttle plasmids pAd-delE1b55kD / 重庆医科大学学报

ABSTRACT

Objective: To construct a replication-competent adenovirus shuttle plasmids named pAd-delE1b55kD,which can transfect the cell of tumor targeted.Methods: delete the E1B55 region of pXC1 and retain a site for Bgl II by such technique of gene recombination as nested PCR;verify the correctness of pAd-delE1b55kD by gel image analysis and sequencing the gene order.Results: The 2279~3326 region of E1B in pXC1 was deleted.The 2024 basic radical C was mutated to T,the 2252 basic radical C to T and the 2261 basic radical G to T.Stop codon TGA?TAG and TAA were inserted.A site for Bgl II was retained in downstream of the stop codon.Conclusion: The replication-competent adenovirus shuttle plasmids named pAd-delE1b55kD is constructed,which provided a carrier for the gene therapy of tumor targeted.