›› 2012, Vol. 39 ›› Issue (4): 42-46.

• 生物技术 • Previous Articles     Next Articles

Secretory Expression of Avian Leukosis Virus Subgroup J gp85 Gene in Pichia pastoris

XU Yan-wei, GAO Hong-lei, GAO Yu-long, LI Kai, GAO Li, QIN Li-ting, QI Xiao-le, WANG Yong-qiang, WANG Xiao-mei   

  1. Division of Avian Infectious Diseases,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute, Chinese Academy Agricultural Sciences,Harbin 150001,China
  • Received:2011-11-05 Revised:1900-01-01 Online:2012-04-20 Published:2012-04-20

Abstract: To express recombinant gp85 protein of avian leukosis virus subgroup J (ALV-J) in P. pastoris, the gp85 gene was amplified and cloned into the pPIC9K vector. The recombinant plasmid pPIC9K-gp85 was identified, linearized with Sal Ⅰ and transformed into P.pastoris strain GS115 by electroporation. Transformants were selected on MD plates then on YPD plates containing G418 and confirmed by PCR. SDS-PAGE assay demonstrated that gp85 protein was expressed and secreted into the culture medium. The expression product amounted to 40 mg/L of culture under optimized condition. The recombinant gp85 protein showed good antigenicity in Western blotting and Dot-ELISA assays. This study laid the foundation for further study of the gp85 protein and the development of the diagnostic methods for ALV-J.

Key words: ALV-J; gp85 gene; Pichia pastoris; secretory expression

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