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采用苜蓿尺蠖核多角休病毒(Autographacalifornicanuclearpolyhedrosisvirus,AcNPV)DNA中多角体蛋白基因启动子及其前后序列作为转移载体,将人β-干扰素(HuIFN-β基因插入转移载体pUAc-5,构建成pAc-IFN-β载体。该质粒DNA与野生型AcNPVDNA经lipofectin共转染秋粘虫(Spodopterafrugiperda)传代细胞(Sf9细胞),利用重组病毒不产生多角体蛋白的特征,进行病毒空斑筛选。用核酸杂交方法鉴定出携带HuIFN-β基因的主组病毒。用重组病毒感染Sf9细胞,可表达rHuIFN-β。在1.0×10 ̄6细胞/ml感染上清中测定rHuIFN-β最高活性为3.0×10 ̄6IU/ml,抗天然HuIFN-β抗体能完全中和rHuIFN-β的抗病毒活性。
The HuIFN-β gene was inserted into the transfer vector pUAc-5 to construct the pAc-5 gene. The promoter of the polyhedrin gene in the DNA of Autographa californica riuclear polyhedrosis virus (AcNPV) IFN-β vector.The plasmid DNA and wild-type AcNPVDNA were co-transfected with Spodoptera frugiperda passage cells (Sf9 cells) by lipofectin, and the virus was screened for virus plaque by using the feature that the recombinant virus did not produce polyhedrin protein Methods The main virus carrying HuIFN-β gene was identified, and rHuIFN-β was expressed in Sf9 cells infected with recombinant virus.The highest activity of rHuIFN-β was detected in 1.0 × 10 6 cells / ml supernatant. 0 × 10 ~ 6IU / ml, anti-natural HuIFN-β antibodies completely neutralize the antiviral activity of rHuIFN-β.