中文摘要：

研究利用Bac-To-Bac杆状病毒表达系统构建含有猪γ-干扰素（porcine interferon-γ,PoIFN-γ）完整开放阅读框的供体质粒pFastBacTM1-PoIFN-γ,转化DH10Bac感受态细胞获得重组穿梭质粒rBacmid-PoIFN-γ,转染sf9昆虫细胞救获表达PoIFN-γ的重组杆状病毒rBac-PoIFN-γ。以抗PoIFN-γ单克隆抗体为一抗进行Western blot、间接免疫荧光（IFA）及间接ELISA检测,结果表明PoIFN-γ在重组杆状病毒rBac-PoIFN-γ感染的昆虫细胞中获得正确表达。抗病毒活性试验显示,重组杆状病毒表达rPoIFN-γ能有效抑制水疱性口炎病毒（VSV）在猪肾细胞系（PK-15）的复制,rBac-PoIFN-γ感染昆虫细胞培养上清抗病毒活性为2×104抑制单位（IU）/mL,其抗病毒活性可以被鼠抗原核表达重组PoIFN-γ免疫血清有效阻断。结果表明rPoIFN-γ在重组杆状病毒rBac-PoIFN-γ在感染昆虫细胞获得有效表达,并具有高效抗病毒活性。

英文摘要：

The full-length porcine interferon gamma（PoIFN-γ） cDNA, including the secretion signal peptide coding region was recloned into honor plasmid pFastBac^TM 1 of Bac-To-Bac Baculovirus Expression System. These recombinant plasmids, pFastBacTM 1-PoIFN-γ, were transformed into DH10Bac host bacteria to get recombinant shuttle plasmids, rBacmid-PoIFN-γ. Recombinant baculovirus, rBac-PoIFN-γ, was generated for expressing PoIFN-γ, by transfecting rBacmid-PoIFN-γ with Cellfectin Reagent into sf9 insect cells. The expression of PoIFN-γ in insect cells was confirmed by Western Blot, indirect immunofluorescence assay and indirect ELISA. The antiviral activity assay shows that PoIFN-γ expressed by the rBac-PoIFN-γ can efficiently inhibit the replication of the recombinant Vesicular Stomatitis Virus expressing green fluorescence protein in PK-15 cells. The antiviral activity of PoIFN-γ can be specifically blocked by anti-PoIFN-γ mouse serum. The antiviral titer of culture supematant of insect cells infected by rBac-PoIFN-7 is 2 × 10^4 IU/mL. The results demonstrat that the rBac-PoIFN-γ can express rPoIFN-γ efficiently and rPoIFN-γ has high antiviral activity.