Arabidopsis RNA immunoprecipitation

被引:51
作者
Terzi, Lionel C. [1 ]
Simpson, Gordon G. [1 ,2 ]
机构
[1] Univ Dundee, Coll Life Sci, Div Plant Sci, Dundee DD2 5EH, Scotland
[2] Scottish Crop Res Inst, Genet Program, Dundee DD2 5DA, Scotland
基金
英国生物技术与生命科学研究理事会;
关键词
Arabidopsis; post-transcriptional; RNA-binding protein; RNA immunoprecipitation; RNA; BINDING PROTEINS; MESSENGER-RNA; FLOWERING TIME; THALIANA; METHYLATION; SPECIFICITY; ENCODES; BRAIN;
D O I
10.1111/j.1365-313X.2009.03859.x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
P>RNA-binding proteins are key regulators of plant gene expression. Consistent with this, the Arabidopsis genome encodes many RNA-binding proteins that are genetically required for normal development and for responding to environmental changes. However, the direct RNA targets and RNA processing events that these RNA-binding proteins control are poorly understood. In order to facilitate the functional characterization of RNA-binding proteins, we have applied the RNA immunoprecipitation assay to Arabidopsis. Working with the U2B '-U2 snRNA interaction as a model experimental system, we show that treatment of intact plants with formaldehyde allows immunocapture of U2 snRNA using antibodies that recognize U2B ' fused to the generic GFP tag. When coupled with recent developments in whole-genome tiling arrays and high-throughput next-generation sequencing, this combination of procedures and technology has the potential to allow systematic functional analysis of plant RNA-binding proteins.
引用
收藏
页码:163 / 168
页数:6
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