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Prokaryotic expression of VP1 gene of footandmouth disease virus and detection of expression product immunogenicity MA
Jingyun, CHEN Feng, CAO Yongchang, ZHOU Qingfeng, BI Yingzuo
(College of Animal Science, South China Agricultural University, Guangzhou 510642,China) |
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Abstract: The recombinant expression vector pBADVP1 was constructed by cloning VP1 gene of footandmouth disease virus into the prokaryotic expression plasmid pBAD/TOPO. The recombinant vector was transformed into the E.coli TOP10 strain. Samples were collected at different induction time after induction with Larabinose .The specificity of the expressed fusion protein was identified with SDSPAGE and Westernblotting. The results showed that the optimal amount of the expressed fusion protein is 26.3% of total bacterial protein after being induced with Larabinose at
0.02g/L concentration for 4 hours. The fusion protein is about 40 ku in size and can effect specific reaction with the antibody. The fusion protein inclusion body was extracted to prepare the oilemulsion vaccine, which was used to immunize cavians subcutaneously. The sera were collected from the cavians and tested for the presence of neutralizing antibody, and the cavians were challenged with FMDV. The results showed that the fusion protein can yield antibodies with high neutralizing activity as well as provide protection against challenge with virulent virus.
Key words: footandmouth disease virus; VP1; expression; immunogenicity |
