Construction of human CEH recombinant adenovirus vector

Objective To construct hCEH recombinant replication deficient adenovirus vector pAd-hCEH amplify in HEK293 cells.Methods hCEH gene was cloned into shuttle vector pAdTrack-CMV to generate a recombinant plasmid pAdTrack-CMV-hCEH,and the resultant plasmid was linearized and was subsequently homogenous recombination with pAdeasy-1 in E.coli BJ5183 cells.Recombinants were screened and were identified by observation of green fluorescent protein(GFP) and PCR.The recombinant plasmid was transfected into HEK293 to package the adenovirus,and the hCEH recombinant adenovirus was observed by fluorescent microscope.Results The hCEH recombinant adenoviral vector was constructed successfully,which was confirmed by restriction enzyme and GFP expression.High titer recombinant adenoviral vector was constructed.Conclusion The hCEH recombinant adenovirus is constructed successfully.