利用MultiBac系统表达有活性鸡γ干扰素及其鉴定  

Expression and Identification of Chicken Interferon-γ with MultiBac System

作　　者：周耀红[1] 冯敏 严会超[1] 孙京臣[1] Zhou Yaohong;Feng Min;Yan Huichao;Sun Jingchen(College of Animal Science,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学动物科学学院,广州510642

出　　处：《蚕业科学》2021年第1期50-58,共9页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(31801030);广州市科技计划项目(202002030218)。

摘　　要：MultiBac系统是一套完善而又强大的真核表达系统,可在昆虫细胞中大量表达高品质的外源蛋白,已经应用于结构生物学研究和药物开发。为了实现鸡γ干扰素(chIFN-γ)在昆虫细胞中的表达,利用零背景转座技术构建重组杆状病毒,通过感染Sf9细胞,表达并且纯化chIFN-γ蛋白。使用制得的多抗血清,在重组杆状病毒感染的Sf9细胞样品中能够检测到大小为17.8 kD的单一条带,表明chIFN-γ成功表达。对纯化得到的重组chIFN-γ蛋白进行生物活性检测,结果显示其抗病毒活性为1.6×10^(4) U/mL,具有良好的生物活性。MultiBac system is a complete and powerful eukaryotic expression system that can produce foreign proteins of high quality and great quantity in insect cells, and has been used in structural biology and drug development. In order to express chIFN-γ protein in insect cells, the zero-background transposition technology was used to construct a recombinant baculovirus, and then chIFN-γ protein was expressed infect Sf9 cells and purified. Then, a single band with a size of 17.8 kD can be detected in the Sf9 cell sample infected with recombinant baculovirus using the prepared polyclonal antiserum, which indicated that chIFN-γ protein was successfully expressed. The purified recombinant chIFN-γ protein was tested for its biological activity and the results showed that it had good biological activity with antiviral activity of 1.6×10^(4) U/mL.

关 键 词：杆状病毒表达系统 鸡干扰素 生物活性 

分 类 号：S852.42[农业科学—基础兽医学]