[Expression of Japanese encephalitis virus E protein in methylotrophic yeast Pichia pastoris].
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.
Wu
Yu-Shui
YS
Department of Microbiology, Fourth Military Medical University, Xi'an 710032,China.
Ren
Jun-Ping
JP
Huang
Qing-Sheng
QS
Ding
Tian-Bing
TB
Zhang
Wei
W
Song
Jian-Hua
JH
Zhu
Zhong-Yong
ZY
Zhang
Hong-Yi
HY
Ma
Wen-Yu
WY
D004591:Electrophoresis, Polyacrylamide Gel; D004664:Encephalitis Virus, Japanese; D015870:Gene Expression; D005822:Genetic Vectors; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics
D011994:Recombinant Proteins