Expression and Immunogenicity Analysis of Spike Glycoprotein of Transmissible Gastroenteritis Virus in Yeast Pichia pastoris
ZHANG Li1, KANG Xue-yan1, ZHANG Zhen-hua1, LI Yong-qing1, GAI Li-li1, JIANG Shi-jin2, ZHANG Pei-jun1
1. Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Beijing 100097, China;
2. College of Veterinary Medicine, Shandong Agricultural University, Taian 271018, China
Abstract：For the good immunogenicity recombinant protein,the Pichia pastoris expression system is used for the production of transmissible gastroenteritis virus recombinant proteins.In this study,the partial spike ( S ) gene (2.2 kb long) of transmissible gastroenteritis virus (TGEV) was amplified using polymerase chain reaction (PCR) with the primers designed according to the complete genome sequence of TGEV in GenBank.The cDNA fragment was sub-cloned into Pichia expression vector pPIC9k (digested with Eco R Ⅰand Not Ⅰ).Recombinant shuttle plasmid DNA,digested with Sal Ⅰ,named pPIC9k- S.The pPIC9k- S was integrated into the genome of the methylotrophic yeast Pichia pastoris GS115 by electroporation.The partial fragment of spike protein (S) of approximately 82 kDa was confirmed by SDS-PAGE and Western-Blotting.The specific antibody against S recombinant protein could be detected by ELISA,the title was 1 : 100,the immunogenicity was good.
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2015, Vol. 30 
