Construction of Human Hepatoma-targeting Recombinant Adenovirus Vector of CD80 and Its Identification

Objective: To construct the hepatoma targeting CD80 gene recombinant adenovirus vector. Methods: The enhancer and promoter of AFP were cloned from the human genomic DNA, and pShuttle-AFP plasmid was generated by replacing the CMV promoter of pShuttle-CMV. CD80 gene was inserted into the downstream of AFP enhancer and promoter, to construct the pShuttle-AFP-CD80 plasmid. After digestion with Pme I, pShuttle-AFP-CD80 was co-transformed into E. coli BJ5183 with adenovirus backbone plasmid, and pAd-AFP-CD80 was constructed by homologous recombination. The recombinant adenovirus DNA was transfected into AD293 cells to prepare adenovirus. After identification and titer of the prepare adenovirus. Results: the cloned AFP enhancer promoterand CD80 gene were identical to those reported in GeneBank. Double-digestion results in line with the expected results. Pac I digestion results are consistent with the purpose of. the pShuttle-AFP-CD80 and pAd-AFP-CD80 were successfully constructed after identification by digestion and PCR. Conclusion: We successfully constructed the human hepatoma targeting CD80 expression adenovirus vector, which laid the foundation for the hepatoma targeting immunotherapy with CD80 molecular.