Prokaryotic Expression and Immunogenicity Analysis of Hemagglutinin-related Outer Membrane Protein from F.necrophorum |
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| Authors: | WU Hong-chao LIU Xiao-ying FENG Er-kai ZHU Yan-zhu WAN Hong-li FENG Zhuo YAN Xi-jun CHEN Li-zhi |
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| Institution: | 1. Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China;2. Jilin Teyan Biological Technology Limited Liability Company, Changchun 130112, China |
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| Abstract: | In order to analyze the immunogenicity of 120 ku hemagglutinin-related outer membrane protein from F.necrophorum,specific primers were designed to amplify the 3 truncated overlapping gene fragments covering the whole open reading frame (ORF) based on the result of antigenic epitope analysis.The gene fragments were ligated into the prokaryotic expression vector to construct pET-28a-p1,pET-32a-p2 and pET-32a-p3,respectively.Then recombinant plasmid was induced expression in E.coli.The SDS-PAGE results showed that the expression of recombinant proteins P1,P2 and P3,molecular mass of the expressed proteins were about 48,59 and 62 ku,respectively.Western blotting indicated that the recombinant proteins could be recognized by antiserum against F.necrophorum from mouse.Humoral immunity response against recombinant proteins was detected in the immunized rabbits.Altogether,these findings suggested that 120 ku hemagglutinin-related outer membrane protein from F.necrophorum had good immunogenicity,and this study provided a reference for further research on genetic engineering subunit vaccine of F.necrophorum. |
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| Keywords: | F necrophorum hemagglutinin-related outer membrane protein prokaryotic expression immunogenicity analysis |
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