title: [Prokaryotic expression, purification of prM of JEV and preparation of monoclonal antibody].
abstract: OBJECTIVE
  To prepare monoclonal antibody (mAb) against prM epitope.
  
  
  METHODS
  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.
  
  
  RESULTS
  mAb against prM epitope of JEV was prepared successfully.
  
  
  CONCLUSIONS
  The obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis.
authors.affiliation: Department of Etiology of Second Military Medical University, Shang hai 200433, China.
authors.forename: Bei-fang
authors.initials: BF
authors.lastname: Ning
authors.forename: Huai-min
authors.initials: HM
authors.lastname: Zhu
authors.forename: Xiao-jun
authors.initials: XJ
authors.lastname: Zhou
authors.forename: Yi
authors.initials: Y
authors.lastname: Cao
authors.forename: Ai-guo
authors.initials: AG
authors.lastname: Zhou
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity; D041702:BALB 3T3 Cells; D002460:Cell Line; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004664:Encephalitis Virus, Japanese / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011387:Prokaryotic Cells / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D014764:Viral Proteins