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Preparation of the Secretory Recombinant ALV-J gp85 Protein Using Pichia pastoris and Its Immunoprotection as Vaccine Antigen Combining with CpG-ODN Adjuvant.

VIRAL IMMUNOLOGY. 2018-07; 
Weifang Jing,* Jinrun Zhou,* Chunyang Wang, Jianhua Qiu, Huijun Guo, and Hongmei Li
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Recombinant Proteins … The protein content was estimated using the dye-binding assay. The lyophilized protein was further concentrated and dissolved in an equilibration buffer (PBS, pH 7.4) and purified by two gel filtration runs using SD200 Nickel column (GenScript USA, Inc., Nanjing, China) … Get A Quote

摘要

This study focuses on preparing the secretory recombinant J subgroup of avian leukosis virus (ALV-J) gp85 protein using Pichia pastoris and evaluating its immunoprotection as vaccine antigen combining with CpGODN adjuvant. The secretory recombinant plasmid pPIC9-gp85 containing ALV-J gp85 gene was designed and was transfected into the genome of P. pastoris (GS115) cells. The recombinant plasmid was expressed under the induction of methanol. The expressed products in the medium of the cells were purified and identified with endoglycosidase digestion assay and western blot mediated with monoclonal antibody (MAb) JE9. The purified product combining with CpG-ODN adjuvant was inoculated intramuscularly into 7-... More

關鍵詞

: J subgroup of avian leukosis virus (ALV-J), secretory protein, Pichia pastoris, immunoprotection,neutralizing antibody
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