Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology · 2003
[Expression of Japanese encephalitis virus E protein in methylotrophic yeast Pichia pastoris].
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Wu Yu-Shui, Ren Jun-Ping, Huang Qing-Sheng, Ding Tian-Bing, Zhang Wei, Song Jian-Hua, Zhu Zhong-Yong, Zhang Hong-Yi, Ma Wen-Yu
Аннотация
AIM: 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.
CONCLUSION: 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.
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.
CONCLUSION: 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.