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Published on: May 6, 2015
[Construction of recombinant adenovirus carrying HBx and mIL-12]
Ya-ou Li1, Ping Chen, Zhi-yong Li
1Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, College of Life Science, Sichuan University, Chengdu 610064, China.
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.
