重组杆状病毒表达牛β干扰素及其生物学活性研究  

Expression of recombinant bovine beta-interferon by baculovirus system and evaluation of its bioactivity

作　　者：陈伟业[1] 肇慧君[2] 王喜军[2] 黄克和[1] 步志高[2] 曹文雁[2] 葛金英[2] 温志远[2] 

机构地区：[1]南京农业大学动物医学院,南京210095 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001

出　　处：《微生物学报》2007年第6期992-996,共5页Acta Microbiologica Sinica

基　　金：国家"973"项目(2005CB523202)~~

摘　　要：本研究构建了表达牛β干扰素的重组杆状病毒,感染sf9细胞后,分别采用免疫荧光和免疫印迹证实了重组BoIFN-β的表达存在于细胞内和培养上清中。采用表达绿色荧光蛋白的水疱性口炎病毒(VSV*GFP)检测分泌到细胞上清中重组BoIFN-β的抗病毒活性,可达到106.0AU/mL。同时重组BoIFN-β还可以激活鸡Mx启动子控制的萤光素酶报告基因的转录表达。综上,本研究采用杆状病毒表达系统,重组牛β干扰素以分泌形式高水平表达,且具有天然Ⅰ型干扰素的生物学活性。In this paper, a recombinant baculovirus containing the ORF of bovine interferon-β （BoIFN-β） gene, rBac- BoIFN-β,was generated to express recombinant BolFN-β （rBoIFN-β） in sf9 insect cells. The expression of rBoIFN-β in rBac-BoIFN-β infecting six） cells and its supernatants was confirmed by indirect immunofluorescence assay and Western blot. The antiviral activity of rBoIFN-β in the supernatant can reach 10^6.0 AU/mL evaluated by the antiviral assay with VSV ＊ GFP that expressed green fluorescence protein, and rBoIFN-β could stimulate the expression of luciferase reporter gene controlled by chicken Mx promoter. All the results showed that rBac-BoIFN-β constructed here could express high level recombinant BoIFN-β in secreted form that had the bioactivity of natural type Ⅰ IFN.

关 键 词：牛β干扰素 抗病毒活性 Mx启动子 重组杆状病毒 

分 类 号：S852.5[农业科学—基础兽医学]