title: [Expression of Japanese encephalitis virus E protein in methylotrophic yeast Pichia pastoris].
abstract: OBJECTIVE
  To express Japanese encephalitis virus (JEV) E protein in methylotrophic yeast Pichia pastoris.
  
  
  METHODS
  The gene coding for JEV E protein was sub-cloned into vector pPIC9k. The constructed plasmid was transformed into yeast SMD1168 by electroporation. The recombinant transformants with a high copy number of the plasmid were selected by using MD plate and G418. The expression of E protein in yeast was induced by the addition of methanol and analyzed by SDS-PAGE and Western blot.
  
  
  RESULTS
  The protein was produced at a yield of 50 mg per litre of culture. Owing to heterogeneous glycosylation, relative molecular mass (M(r)) of the expressed E protein was sized about 113 000 and 78 000.
  
  
  CONCLUSIONS
  JEV E protein was expressed successfully in yeast Pichia pastoris, which should be useful for the production of diagnostic reagents and genetically engineered vaccine of JEV.
authors.affiliation: Department of Microbiology, Fourth Military Medical University, Xi'an 710032,China.
authors.forename: Yu-Shui
authors.initials: YS
authors.lastname: Wu
authors.forename: Jun-Ping
authors.initials: JP
authors.lastname: Ren
authors.forename: Qing-Sheng
authors.initials: QS
authors.lastname: Huang
authors.forename: Tian-Bing
authors.initials: TB
authors.lastname: Ding
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Jian-Hua
authors.initials: JH
authors.lastname: Song
authors.forename: Zhong-Yong
authors.initials: ZY
authors.lastname: Zhu
authors.forename: Hong-Yi
authors.initials: HY
authors.lastname: Zhang
authors.forename: Wen-Yu
authors.initials: WY
authors.lastname: Ma
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004664:Encephalitis Virus, Japanese; D015870:Gene Expression; D005822:Genetic Vectors; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D011994:Recombinant Proteins