Construction of recombinant adenoviral vector co-expressing interleukin-7 and enhanced green fluorescent protein

Objective:To construct the adenoviral vector co-expressing interleukin-7(IL-7) and enhanced green fluorescent protein(EGFP),to lay a experimental foundation for further study on the infection into stem cell.Methods: The target gene IL-7 was cloned into the shuttle plasmid expressed the report gene EGFP.Then the recombinant shuttle plasmid was transformed into Dh5a bacteria to recombine with backbone vector pAdxsi.Next,the plasmid pAd-EGFP-mIL7 was amplified in H293 cells and purifired,then the viral titer was determined.Results: The recombinanted shuttle plasmid pShuttle-EGFP-mIL7 digested with restriction endonucleases was confirmed by two products which length were respectively about 0.5kb and 5.1kb;the recombinanted plasmid pAdxsi-EGFP-mIL7 digested with restriction endonucleases was confirmed by seven products which length were respectively about 14K,11.8K,3.1kb,2.66kb,2.47K,1.45K and 0.6K;recombinant adenoviral amplifired with titer of 2×1010pfu/ml.Conclusion: The recombinant adenoviral vector pAdxsi-EGFP-mIL7 was successfully constructed.