Miyoshi Keita, Ogino Akiyo, Siomi Mikiko C, Siomi Haruhiko
Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.
Methods Mol Biol. 2013;1010:111-21. doi: 10.1007/978-1-62703-411-1_8.
Fragile X syndrome results from the lack of FMR1 expression. To understand how the lack of FMR1 function leads to the syndrome, we are studying the Drosophila FMR1 related protein (dFMR1). We performed affinity purification of dFMR1-associated complexes from cultured Drosophila S2 cells and found that dFMR1 associates with a key component of RNA interference, AGO2. Our finding suggests cross talk between the fragile X syndrome protein and RNA interference. In this chapter, we describe a tandem affinity purification method to isolate the protein components and small RNAs in the dFMR1-associated complexes.
脆性X综合征是由于FMR1基因不表达所致。为了解FMR1功能缺失如何导致该综合征，我们正在研究果蝇FMR1相关蛋白（dFMR1）。我们从培养的果蝇S2细胞中对与dFMR1相关的复合物进行了亲和纯化，发现dFMR1与RNA干扰的关键成分AGO2相关联。我们的发现表明脆性X综合征蛋白与RNA干扰之间存在相互作用。在本章中，我们描述了一种串联亲和纯化方法，用于分离与dFMR1相关复合物中的蛋白质成分和小RNA。
