Construction and expression of multiple epitopes DNA vaccines of influenza A virus H3579 subtypes
Objective To construct DNA vaccines which express influenza A viruses multiple epitopes, and to study the bioactivity of the expression products in BHK cells. Methods Selecting the main antigen (HA, NA, NP, M) dominant epitopes of influenza A viruses, a sequence was synthesized containing multiple epitopes of influenza A viruses. A box co-expressing H7H5HA and Epi gene was contructed by inserting the Epi gene in H7H5HA ORF. The box was then cloned into pVAX1. To detect the mRNA of the gene of interest the whole RNA of the BHK cells transfected with the reconstructed plasmids pVAX1, pVAX1-H7HA-Epi-H5HA by liposome method were extracted. 1FA was also utilized to detect the bioactivity of the expression products. Results The gene of interest was identified by RT-PCR, and the antibody specificity of expression products was detected by IFA. Conclusion The reconstructed plasmid could express product efficiently, and the product can specially rec- ognized by the corresponding antibodies. This proved the reconstructed eukaryotic expression vector pVAX1-H7HA- Epi-H5HA might be a candidate vaccine strains for influenza A viruses.