Construction of Human FRNK Recombinant Adenovirus Vector

Objective: To construct an hFRNK adenovirus vector,pAdhFRNK.Methods: Human FRNK was cloned into shuttle vector pAdTrack-CMV to generate a recombinant plasmid pAdTrack-CMV-hFRNK,and the resultant plasmid was linearized by Pme Ⅰ enzymatic digestion and was subsequently electroporated into E.coli BJ5183 cells that had been electroporated adenovirus backbone plasmid pAdEasy-1.Recombinants were screened by Kanamycin and were identified by observation of green fluorescent protein(GFP) expression and PCR.The Pac Ⅰ-linearized recombinant plasmid was transfected into HEK293 cells to package the adenovirus,and the hFRNK recombinant adenovirus was observed by fluorescent microscope.Results: The hFRNK recombinant adenoviral vector was constructed successfully,which was confirmed by restriction enzyme and GFP expression.Conclusion: The human FRNK recombinant adenovirus was constructed successfully,and provided a sound base for gene therapy.