Construction of recombinant adenovirus carrying human DeltaNp73α cDNA with Adeasy system

Objective To construct recombinant adenovirus carrying human DeltaNp73α cDNA.Methods Human DeltaNp73α cDNA was cloned into adenovirus transfer vector pAdTrack-CMV.The obtained plasmid,pAdTrack-CMV-DeltaNp73α,was then linearized with PmeⅠ and transformed into E.coli.strain BJ5183 containing pAdeasy-1,the viral DNA plasmid,for homologous recombination.The adenoviral recombinant was then cleaved with PacⅠ and transfected into 293T cells to produce viral particles.The virus was then transfected into dentritic cells,and the expression of DeltaNp73α was proved by GFP expression and Western blotting.Results Recombinants were confirmed right by restriction enzyme analysis,PCR on viral lysate and expression of green fluorscence protein.A band of 67×103 was observed by Western blotting as expected.Conclusion The recombinant adenovirus carrying human DeltaNp73α cDNA is successfully constructed,paving the way for further research about its high expression in interested cells.