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RNA干扰被认为是转录后基因沉默的一种机制。RNA干扰通过小干扰RNA特异性降解目标mRNA来沉默基因表达。本文以烟草花叶病毒126kD蛋白为靶蛋白,在原生质体水平上研究了小干扰RNA对病毒侵染的干扰和抑制作用。ELISA和Northern杂交的实验结果表明,共转染小干扰RNA和TMV的原生质体内检测到较低的病毒含量。在枯斑寄主上,叶片接种小干扰RNA和TMV共转染原生质体后,与对照叶片相比,仅有很少量的病斑产生。这说明,小干扰RNA能够在原生质体水平对病毒起到干扰和抑制作用。因此认为,烟草原生质体系统有利于快速和定量分析小干扰RNA介导对植物病毒的抑制作用。
RNA interference is considered as a mechanism of post-transcriptional gene silencing. RNA interference silences gene expression by specific degradation of target mRNAs by small interfering RNAs. In this paper, tobacco mosaic virus 126kD protein as the target protein at the level of protoplasts to study the interference of small interfering RNA on viral infection and inhibition. The experimental results of ELISA and Northern blot showed that a lower virus content was detected in protoplasts co-transfected with small interfering RNA and TMV. On the blight host, only a very small amount of lesions were produced compared to the control leaves after the leaf was inoculated with small interfering RNA and TMV co-transfected protoplasts. This shows that small interfering RNA can interfere with and inhibit the virus at the level of protoplast. Therefore, tobacco protoplast system is conducive to rapid and quantitative analysis of small interfering RNA mediated inhibition of plant viruses.