BACKGROUND: Encoded by the UL49 gene of herpes simplex virus type 1 (HSV-1), VP22 is an alkaline protein. With the protein transduction domain (PTD), VP22 can mediate a transmembrane transduction of VP22-linked protein or DNA from the cells in which it is synthesized endogenously to adjacent cells, which shows advantage in gene targeting prevention.
OBJECTIVE: To construct a recombinant adenovirus based vaccine in order to express Vp22 of HSV-1 and the VP1, the main neutralizing antigen of coxsackievirus B3 (CV B3), and to observe the expression of exogenous gene in HEK293 cells. 
METHODS: The DNA fragments of HSV-1 VP22 and CVB3 VP1 were amplified by PCR and linked by linker to produce VP22-L-VP1. The VP22-L-VP1 coding sequence was cloned to the pAdTrack-CMV plasmid to construct AdTrack-CMV/VP22-L-VP1. Then AdTrack-CMV/VP22-L-VP1 was transformed into Ecoli.Bj5183 carrying backbone plasmid pAdEasy-1 to produce the recombinant adenovirus plasmid pAd/VP22-L-VP1. HEK293 cells were transfected with pAd/VP22-L-VP1 to produce recombinant adenovirus rAd/VP22-L-VP1. The virus amplification and titration was preformed on HEK293 cells and the exogenous gene expression was detected by Western blot.
RESULTS AND CONCLUSION: The titer of recombinant adenovirus rAd/VP22-L-VP1 of passage 4 was 6.77×107 pfu/ml. And the expression of VP22-L-VP1 was verified on HEK293 cells by Western blotting. Recombinant adenovirus rAd/VP22-L-VP1 was generated successfully, which laid a foundation for further study on CVB3VP1 gene vaccine.