RNA干扰抑制猪繁殖与呼吸综合症病毒N基因表达及病毒复制  

RNA interference mediated inhibition of PRRSV N gene expressions and virus replication in Marc145 cells

作　　者：黄良宗[1] 钟科阳[1] 王淑敏[1] 司兴奎[1] 马春全[1] 顾万军[1] 

机构地区：[1]佛山科学技术学院广东省高校预防兽医学重点实验室,广东佛山528231

出　　处：《广东农业科学》2011年第11期148-150,F0003,共4页Guangdong Agricultural Sciences

基　　金：广东省自然科学基金(9152800001000017)

摘　　要：根据siRNA设计原则,选取了PRRSV N基因上的保守序列作为可能的干扰位点,设计、合成了3个siRNAs,分别将其克隆到pSIREN-Shuttle中,获得3个siRNA重组表达质粒:pShuttle-N1,pShuttle-N2和pShuttle-N3。将siRNA重组表达质粒与融合表达质粒pEGFP-ORF7共转染Marc145细胞,并通过荧光显微镜观察。结果显示,siRNA重组表达质粒能显著抑制PRRSV N基因的表达。在转染总剂量(0.7μg/孔)固定的条件下,分别将不同组合的siRNA表达质粒共转染Marc145细胞,各组合转染孔对PRRSV复制的抑制作用没有明显差异,其中3种siRNA表达质粒共转染对PRRSV复制的抑制最好,证实siRNA对病毒复制作用具有相加性。According to the design principle （~f the siRNA, three siRNAs directed against PRRSV N gene were synthesized and then cloned into the plasmid pSIREN-Shuttle. Three recombinant siRNA expression plaso, lds pShuttle-N 1, pShuttle-N2 and pShuttle-N3 were constructed successfully, pEGFP-CI plasmid was transfected the Marc145 cells lines to study the transfection efficiency of siRNA plasmids. In order to detect the inhibition effect of the target gene, recombinant siRNA expression plasmid pShuttle-N1, pShuttle-N2 and pShuttle-N3 were co-transfected Mare145 cells respectively with recombinant expression plasmid pEGFP-ORF7. The green fluorescence was observed under UV light. The results showed that recombinant siRNA could dramatically inhibit the expression of PRRSV related gene. Different groups of siRNA expression plasmids were co-transfected Marc145 cells with same dose. The result showed that there was no significant difference between co-transfection in viral inhibitory efficiency, but the inhibitory effect of three siRNA expression plasmid co-transfection was better than the both.

关 键 词：猪繁殖与呼吸综合征病毒(PRRSV) RNA干扰 病毒复制 

分 类 号：Q785[生物学—分子生物学]