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標題: Secreted expression of the VP2 protein of very virulent infectious bursal disease virus in the methylotrophic yeast Pichia pastoris
作者: Wu, P.C.
Su, H.Y.
Lee, L.H.
Lin, D.T.
Yen, P.C.
Liu, H.J.
關鍵字: methylotrophic yeast Pichia pastoris;very virulent infectious bursal;disease virus (vvIBDV);Mut(+) phenotype;double-stranded-rna;escherichia-coli;genomic segment;2 serotypes;sequence;strains;protection;identification;vaccination;antibodies
Project: Journal of Virological Methods
期刊/報告no:: Journal of Virological Methods, Volume 123, Issue 2, Page(s) 221-225.
The VP2-encoding gene of very virulent infectious bursa] disease virus (vvIBDV) was amplified using reverse transcription (RT)-polymerase chain reaction (PCR) and inserted into pPICZalphaA vector. Recombinant plasmid DNA was integrated into the chromosome of the transformed Pichia pastoris by electroporation and expressed protein identified by SDS-PAGE and Western blotting. High-level secreted expression was performed by determining the Mut(+) phenotype and secreting multi-copy integrants in the recombinant yeast. A recombinant protein of approximately 67 kDa was secreted into the supernatant from the yeast when induced with methanol. The expressed supernatant was bound with chicken anti-IBDV polyclonal antibodies. Western blotting with antibodies against vvIBDV indicated that the recombinant VP2 protein retained its antigenicity. High-level production (10mg/100m]) of the recombinant VP2 protein indicated that P pastoris was an efficent expression system for vvIBDV VP2 protein. (C) 2004 Elsevier B.V. All rights reserved.
ISSN: 0166-0934
DOI: 10.1016/j.jviromet.2004.10.002
Appears in Collections:分子生物學研究所

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