鸡γ-干扰素在毕赤酵母中的表达及其生物活性检测  

Biological Activity of the Recombinant Chicken Interferon Gama Expressed in Pichia pastoris

作　　者：李亚玲[1] 秦乐[1] 张小荣[1] 吴艳涛[1] 

机构地区：[1]扬州大学兽医学院农业部禽用生物制剂创制重点实验室,江苏扬州225009

出　　处：《中国兽医杂志》2015年第3期34-35,39,共3页Chinese Journal of Veterinary Medicine

基　　金：现代农业产业技术体系专项资金(No.CARS-41-K08);江苏高校优势学科建设工程资助项目

摘　　要：将鸡γ-干扰素(Ch IFN-γ)基因克隆至p PICZαA,以线性化质粒p PICZαA-IFN-γ电转化酵母GS115细胞,筛选出表达重组Ch IFN-γ(r Ch IFN-γ)的酵母菌株。r Ch IFN-γ通过离子交换层析纯化后,经水泡性口炎病毒(VSV)检测其抗病毒活性为10 240 IU/m L。以r Ch IFN-γ与H9亚型禽流感灭活疫苗联合免疫SPF鸡,使用r Ch IFN-γ试验鸡的血清HI抗体水平显著高于未使用r Ch IFN-γ试验鸡(P<0.05);用H9亚型禽流感病毒攻击后,使用r Ch IFN-γ试验鸡排毒率显著低于未使用r Ch IFN-γ试验鸡(P<0.05),说明r Ch IFN-γ具有佐剂的活性。The nucleotide sequence of ChlFN-γ mature protein was cloned into pPICZotA to construct recombinant plasmid pPICZαA-IFN-γ. After linearized, the recombinant plasmid was then transformed into the P. pastoris GS115 by electroporation. Recombinant strains were obtained and induced to express IFN-% After purified by DEAE anion exchange chromatography, the antiviral activity of rChIFN-γ reached 10 240 IU/mL tested by VSV system. To evaluate the adjuvant effect of rChIFN-γ, SPF chickens were immunized with rChIFN-γ and H9 subtype avian influenza virus （AIV）inactivated vaccine. The results demonstrated that the HI antibody titers of chickens immunized with vaccine and rChIFN-γ were significantly higher than chickens without rChIFN-γ/（P〈 0.05）. After challenged with H9N2 AIV, the virus shedding rates of chickens immunized with vaccine and rchlFN-γ were significantly lower than chickens without rChIFN-γ（P〈0.05）.

关 键 词：鸡γ-干扰素 毕赤酵母 表达 纯化 佐剂 

分 类 号：S852.4[农业科学—基础兽医学]