Citation: ZHANG Xue-han, HE Kong-wang, GUO Rong-li, NI Yan-fang, WANG Fang, YU Zheng-yu. Cloning and Expression of E Gene of Japanese Encephalitis Virus and Preliminary application .VIROLOGICA SINICA, 2005, 20(5) : 494-497.

Cloning and Expression of E Gene of Japanese Encephalitis Virus and Preliminary application

  • Available online: 20 October 2005
  • The whole cDNA of E gene was amplified by RT-PCR from JEV strain SA14-14,and cloned into the pMD18-T vector.The fragment was identified by restriction enzymes digestion with EcoR I and Xho I,cloned into the pET32a(+) vector.The recombinant plasmid was transformed into BL21 and the recombinant bacteria was induced by optimal concentration of IPTG.SDS-PAGE and Western blotting were performed to detect the E fusion protein.Our result indicates the molecular weight of E protein is of E protein 73kDa and its antigenicity is good.Indirect ELISA procedure has been developed with the purified E protein as antigen for detection of JEV.

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    Cloning and Expression of E Gene of Japanese Encephalitis Virus and Preliminary application

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    Abstract: The whole cDNA of E gene was amplified by RT-PCR from JEV strain SA14-14,and cloned into the pMD18-T vector.The fragment was identified by restriction enzymes digestion with EcoR I and Xho I,cloned into the pET32a(+) vector.The recombinant plasmid was transformed into BL21 and the recombinant bacteria was induced by optimal concentration of IPTG.SDS-PAGE and Western blotting were performed to detect the E fusion protein.Our result indicates the molecular weight of E protein is of E protein 73kDa and its antigenicity is good.Indirect ELISA procedure has been developed with the purified E protein as antigen for detection of JEV.

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