Avian reovirus σC gene was virus adhesion proleins,and σNS gene had the activity of binding single strand RNA. According to the siRNA target sequences,siRNA templates were designed, synthesized and then cloned into the shRNA expression vector pSilencer-CMV 4.1 neo. ShRNA vector C1, C2, C3 for σC gene and shRNA vectora for the σNS gene NS1, NS2, NS3 were constructed separately. ShRNA vectors of σC and σNS gene and negative control shRNA were co-transfected into DF-1 cells with the eukaryotic expression vector pECFP-σC and pEGFP-σNS, respectively. Fluorescence microscopy results indicated that the constructed 6 shRNA fragments could inhibit the expression of fusion protein in some degrees. Real-time PCR tests demonstrated that C3 and NS1 had the best interference effect to the viral replication in vitro.%禽呼肠孤病毒σC基因是病毒粘附蛋白,σNS基因具有结合单链RNA活性.根据siRNA靶序列设计原则,设计并合成siRNA模板并克隆到shRNA表达载体pSilencer-CMV 4.1 neo.分别构建了针对σC基因的shRNA载体C1、C2、C3和针对σNS基因的shRNA载体NS1、NS2、NS3.将构建的shRNA载体和阴性对照分别与表达σC和σNS基因的融合蛋白的真核表达载体pEG-FP-σC及pEGFP-σNS共转染DF-1细胞.荧光显微镜观察结果表明,6个shRNA片段不同程度地抑制各自融合蛋白的表达.Realtime PCR检测结果表明,C3和NS1体外干扰病毒复制的效果最佳.
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