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机构地区:[1]南京农业大学动物医学院
出 处:《中国病毒学》2005年第3期272-276,共5页Virologica Sinica
摘 要:为探讨短的双链RNA(siRNA)对鸡传染性支气管炎病毒(IBV)增殖的干扰作用,利用软件设计siRNA1280个,75%位于Pol基因内。通过同源比较和保守性分析,筛选到针对Pol、M、N基因的12个siRNA(每个基因3~4个)作为后选目的片段,分别在Vero细胞、9日龄SPF鸡胚上进行基因干扰试验。结果,来自Pol、N靶序列的2个siRNA在Vero细胞上及鸡胚上均对IBV增殖产生明显的干扰作用,并与siRNA剂量有一定相关性,依赖于与mRNA互补的负链siRNA存在。本研究首次证实IBV增殖过程中存在siRNA干扰现象,为利用RNA干扰(RNAi)技术控制IBV提供了新手段。The short interfering RNAs(siRNA) specific for conserved regions of avian Infectious bronchihitis virus(IBV) genome were designed using the software online. The siRNA had the potency of inhibiting IBV production in both cell lines and embryonic chicken eggs. 12 siRNAs from polymerase gene (pol), M and N gene were used for the experiments. N1221 from N gene and Pol50 from pol gene were shown to have an obvious RNAi effect on IBV production in Vero cells and 9 day-old SPF chicken embryos. The RNAi effect depending on the presence of a functional antisense strand of the siRNA duplex showed the positive relation with transfection dosage. It suggested that viral RNA or mRNA was the target of RNAi. This report firstly revealed RNAi phenomenon of IBV replication procedure in Vero cells and 9 day-old SPF chicken embryos.
关 键 词:鸡传染性支气管炎病毒 SIRNA RNAI VERO细胞 SPF鸡胚
分 类 号:S852.65[农业科学—基础兽医学]
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