Construction of adenovirus vector carrying apoptin gene and endostatin gene

Objective To construct recombinant adenovirus vector carrying apoptin (VP3) gene and endostatin gene. Methods vp3 gene and endostatin gene were cloned into adenovirus shuttle plasmid pDC316. Then the recombinant shuttle plasmid was cotransformed into HEK293 cells with adenovirus backbone plasmid pBHGloxE1 to obtain recombinant adenovirus particles (Ad-vp3-IRES-sEndo-his). The adenovirus particles was screened and confirmed by PCR and RT-PCR, and the correct adenovirus clones screened were amplified in large scale and purified. The viral particle titration such as tissue culture infectious dose 50 (TCID50) and virus particles (VP) was detected. Results PCR and RT-PCR test indicated that apoptin gene and endostatin gene were integrated into the adenoviral genome correctly. Conclusion The recombinant adenovirus vector carrying VP3 gene and endostatin gene was successfully constructed, which qualifies for use in cell and animal experiments.