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Chinese Journal of Experimental and Clinical Virology ; (6): 65-67, 2008.
Article in Chinese | WPRIM | ID: wpr-254138

ABSTRACT

<p><b>OBJECTIVE</b>To prepare monoclonal antibody (mAb) against prM epitope.</p><p><b>METHODS</b>The gene encoding prM was isolated using RT-PCR from brain of JEV infected mouse and cloned into prokaryotic expression vector pET-32a. Recombinant plasmid was transformed into E.coli BL21/DE3/LysS, then the transformed cells were expressed with the induction of IPTG. The expression and purification of the prM protein was analyzed by SDS-PAGE. The BALB/c mice were immunized with purified prM protein. Hybridoma cell lines secreting monoclonal antibodies against prM were established after cell fusion of mouse splenic cell and P3-X63-Ag8.653 cells. The specificity of mAb was identified by ELISA, Western Blot and Immunohistochemistry assay.</p><p><b>RESULTS</b>mAb against prM epitope of JEV was prepared successfully.</p><p><b>CONCLUSION</b>The obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis.</p>


Subject(s)
Animals , Mice , Antibodies, Monoclonal , Allergy and Immunology , Antibody Specificity , BALB 3T3 Cells , Cell Line , Cloning, Molecular , Electrophoresis, Polyacrylamide Gel , Encephalitis Virus, Japanese , Genetics , Allergy and Immunology , Epitopes , Allergy and Immunology , Escherichia coli , Genetics , Plasmids , Genetics , Metabolism , Prokaryotic Cells , Metabolism , Sequence Analysis, DNA , Viral Proteins , Genetics , Allergy and Immunology
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