【摘要】 为研究RNA干扰对捻转血矛线虫Hc38基因转录的抑制作用,针对Hc38基因序列设计并合成特异性dsRNA、3对siRNAs(siRNA1,siRNA2,siRNA3),采用浸泡方式分别将dsRNA、siRNAs导入捻转血矛线虫感染性L3期幼虫体内,利用实时荧光定量PCR分析Hc38基因在L3期幼虫中的相对含量以确定抑制效果。通过体外转录成功获得高浓度和高纯度的dsRNA,L3期幼虫经浸泡处理3d后,siRNAs组Hc38基因的相对含量分别为(14.93±1.75)%、(26.34±2.91)%和(15.86±2.54)%,dsRNA组为(32.27±1.50)%,均显著低于对照组。表明通过浸泡法导入的dsRNA和siRNAs均能有效抑制Hc38基因的转录,从而为捻转血矛线虫及其他寄生线虫的基因功能研究提供了一种新的方法。更多还原

【Abstract】 To study RNA interference effect on suppression of transcription of Haemonchus contortus Hc38 gene,dsRNA and siRNAs(siRNA1,siRNA2,siRNA3) were designed and synthesized specifically according to the Hc38 gene sequence,and transferred into H.contortus L3 by soaking.Interference effect was examined by real-time PCR.The dsRNA with high concentration and purity was synthesized successfully by transcription in vitro.After soaking for 3 days,the relative content of Hc38 genes in the siRNAs groups was(14.93±1.75)%,(26.34±2.91)% and(15.86±2.54)%,while the dsRNA group was(32.27±1.50)%,significantly lower than that of control group.Results showed that both dsRNA and siRNAs can both inhibit the transcription of Hc38 gene effectively by soaking,and it may provide a new method for the studies of gene function of H.contortus and other parasitic nematodes.更多还原