[Construction of recombinant adenovirus carrying HBx and mIL-12].

ABSTRACT

OBJECTIVE: To construct a recombinant adenovirus Ad-HBx-mlL-12 carrying HBx and mIL-12. METHODS: HBx and mIL-12 were cloned into the shuttle plasmid pAdenoVator-CMV5 and confirmed by means of enzymatic manipulation. After linearization by EcoR I digestion, the recombinant shuttle plasmid pAdV-HBx-mIL-12 was transformed into competent BJ5183 germs with the adenoviral backbone plasmid pAdenoVator deltaE1/E3 and then homologically recombined to obtain the recombinant adenovirus plasmid. After confirmation, the recombinant adenovirus plasmid pAd-HBx-mIL-12 was linearized with Pac I digestion and transfected into 293 cells via liposome, and then adenovirus package and amplification were performed. RESULTS: It was confirmed that the recombinant adenovirus Ad-HBx-mIL-12 had been successfully constructed and both HBx and mIL-12 were expressed in 293 cells. CONCLUSION: The recombinant adenovirus carrying HBx and mIL-12 has been successfully constructed, which lays a foundation for the further study of antitumor mechanism and gene therapy.