ObjectiveTo construct a 5/11 chimeric adenovirus harboring RGD-4C,and observe its transfection efficiency after transfecting it into hepatocarcinoma cell lines HepG2,BEL-7404 and BJ cells.MethodsRGD-4C was inserted into the HI loop region of 5/11 chimeric adenovirus fiber gene and the insertion outcome was confirmed by enzyme digestion and PCR analysis.The confirmed plasmid was co-transfected into E.coli BJ5183 together with adenoviral backbone plasmid pPE3 to produce recombinant plasmid by homologous recombination.Recombinants were selected and co-transfected into 293 cell line with PDC328-EF1-EGFP to produce recombinant adenovirus.The recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified.The virus titer of recombinant adenovirus was determined.AD11-RGD-4C-EGFP was used to infect HepG2,BEL-7404,and BJ cell lines,and their transduction efficiency was determined by fluorescence microscope.ResultsA 1 123 bp target gene fragments was obtained by PCR from the recombinant adenovirus; meanwhile,we prepared high titer recombinant adenovirus,with the titer being 1.3×1010pfu/ml after purification.At 10 MOI,the infection efficiency of recombinant adenovirus was much higher than control adenovirus(AD11-EGFP) after 48 h infection.ConclusionWe have successfully constructed a chimeric adenovirus harboring RGD-4C,which paves a way for enhancing the infection efficiency of adenovirus in bio-therapy.