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通过单链DNA亲和柱色谱法分级分离后,水稻干种子中RNA结合蛋白的蛋白质组学分析。

Proteomic analysis of RNA-binding proteins in dry seeds of rice after fractionation by ssDNA affinity column chromatography.

作者信息

Masaki Shunpei, Yamada Tetsuya, Hirasawa Tadashi, Todaka Daisuke, Kanekatsu Motoki

机构信息

Department of Plant Production United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Fuchu, 183-8509, Tokyo, Japan.

出版信息

Biotechnol Lett. 2008 May;30(5):955-60. doi: 10.1007/s10529-007-9619-8. Epub 2007 Dec 18.

Abstract

In proteomic analysis, one of the major limitations is the detection of low-abundance proteins. To detect low-abundance RNA-binding proteins in mature dry seeds of rice, fractionation by single stranded DNA (ssDNA) affinity column chromatography was carried out before analysis by two-dimensional gel electrophoresis (2-DE). Proteomic analysis of the ssDNA-binding fraction revealed the existence of three types of RNA-binding proteins, including a K homology (KH) domain containing protein, a putative RNA-binding protein and a glycine-rich RNA-binding protein, in mature seeds. In addition, decreases in the putative RNA-binding protein and glycine-rich RNA-binding protein after absorbing water in seeds appear to be associated with seed germination.

摘要

在蛋白质组学分析中,一个主要限制是低丰度蛋白质的检测。为了检测水稻成熟干燥种子中的低丰度RNA结合蛋白,在通过二维凝胶电泳(2-DE)分析之前,进行了单链DNA(ssDNA)亲和柱色谱分离。对ssDNA结合部分的蛋白质组学分析揭示了成熟种子中存在三种类型的RNA结合蛋白,包括一种含K同源(KH)结构域的蛋白质、一种假定的RNA结合蛋白和一种富含甘氨酸的RNA结合蛋白。此外,种子吸水后假定的RNA结合蛋白和富含甘氨酸的RNA结合蛋白的减少似乎与种子萌发有关。

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