Prokaryotic Expression and Antigenicity Analysis of Porcine Japenese Encephalitis Virus Envelope Protein Domain Ⅲ |
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Authors: | HOU Hua-yan DING Ling-ling LV Mao-jie YANG Bao-shou |
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Institution: | 1. Ringpu Biological Research Institute, Tianjin Ringpu Biotechnology Co., Ltd., Tianjin 300308, China;2. Key Laboratory of Biogenic Veterinary Drugs Innovation, Ministry of Agriculture, Tianjin 300308, China |
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Abstract: | In order to analyze the antigenicity of porcine Japanese encephalitis virus (JEV) E protein domain Ⅲ, which was expressed by pET-28a vector with His-tag and purified through Ni-NTA, the BALB/c mice were immunized with the purified protein.We identified the antigenicity of domain Ⅲ of E protein and the anti-mice and anti-porcine JEV E Ⅲ protein specific antibody titers by SDS-PAGE, Western blotting, indirect ELISA and IFA.SDS-PAGE results showed the expressed target protein existed mainly in the form of inclusion body.Western blotting, ELISA test results showed that the protein had good reactivity with anti-serum.The mice immunized with the purified JEV E Ⅲ protein generated 1×105 anti-JEV E Ⅲ protein specific antibody titers by ELISA, and the porcine immunized with the porcine JEV generated 5.1×104 anti-JEV specific antibody titers.The IFA results showed that JEV E Ⅲ protein anti-serum could identify JEV antigen.The above results showed that the recombinant JEV E Ⅲ had good antigenicity.These results provided important basis for development of diagnostic antigen for JEV. |
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Keywords: | porcine Japanese encephalitis virus E protein domain Ⅲ diagnostic antigen |
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