Ye Xuecheng, Liu Qinghua
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Methods Mol Biol. 2008;442:11-27. doi: 10.1007/978-1-59745-191-8_2.
RNA interference (RNAi) is a form of posttranscriptional gene silencing mediated by microRNA (miRNA) and small interfering RNA (siRNA). In Drosophila melanogaster, the RNase III enzymes Dicer-1 and Dicer-2 generate miRNA and siRNA, respectively. We describe the methods for the expression, purification, and analysis of recombinant Dicer-1 and Dicer-2 enzymes. Our studies demonstrate that Dicer-1 and Dicer-2 display different substrate specificities and ATP requirements.
RNA干扰(RNAi)是一种由微小RNA(miRNA)和小干扰RNA(siRNA)介导的转录后基因沉默形式。在黑腹果蝇中,核糖核酸酶III(RNase III)家族的Dicer-1和Dicer-2酶分别产生miRNA和siRNA。我们描述了重组Dicer-1和Dicer-2酶的表达、纯化及分析方法。我们的研究表明,Dicer-1和Dicer-2表现出不同的底物特异性和ATP需求。