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

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10^6.0AU/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。

Abstract:

In this paper，a recombinant baculovirus containing the ORF of bovine interferon-β (BoIFN-β) gene，rBac-BoIFN-β，was generated to express recombinant BoIFN-β (rBoIFN-β) in sf9 insect cells. The expression of rBoIFN-β in rBac-BoIFN-β infecting sf9 cells and its supernatants was confirmed by indirect immunofluorescence assay and Western blot. The antiviral activity of rBoIFN-β in the supernatant can reach 10^6.0AU/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.