用于全基因组芯片分析的内源性蛋白质的表位标记
Epitope tagging of endogenous proteins for genome-wide ChIP-chip studies.
作者信息
Zhang Xiaodong, Guo Chunguang, Chen Yueting, Shulha Hennady P, Schnetz Michael P, LaFramboise Thomas, Bartels Cynthia F, Markowitz Sanford, Weng Zhiping, Scacheri Peter C, Wang Zhenghe
机构信息
Department of Genetics and Case Comprehensive Cancer Center, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio 44106, USA.
出版信息
Nat Methods. 2008 Feb;5(2):163-5. doi: 10.1038/nmeth1170. Epub 2008 Jan 6.
Abstract
We developed a strategy to introduce epitope tag-encoding DNA into endogenous loci by homologous recombination-mediated 'knock-in'. The tagging method is straightforward, can be applied to many loci and several human somatic cell lines, and can facilitate many functional analyses including western blot, immunoprecipitation, immunofluorescence and chromatin immunoprecipitation-microarray (ChIP-chip). The knock-in approach provides a general solution for the study of proteins to which antibodies are substandard or not available.
摘要
我们开发了一种策略，通过同源重组介导的“敲入”将编码表位标签的DNA引入内源性基因座。这种标记方法简单直接，可应用于许多基因座和多种人类体细胞系，并且能够促进包括蛋白质免疫印迹、免疫沉淀、免疫荧光以及染色质免疫沉淀微阵列（ChIP芯片）在内的多种功能分析。敲入方法为研究缺乏优质抗体或没有可用抗体的蛋白质提供了一个通用的解决方案。
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