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利用染色质免疫沉淀技术绘制酿酒酵母中的共转录mRNA加工图谱。

Using chromatin immunoprecipitation to map cotranscriptional mRNA processing in Saccharomyces cerevisiae.

作者信息

Keogh Michael-Christopher, Buratowski Stephen

机构信息

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.

出版信息

Methods Mol Biol. 2004;257:1-16. doi: 10.1385/1-59259-750-5:001.

Abstract

The chromatin immunoprecipitation (ChIP) technique has been used to determine where and under what conditions DNA binding proteins associate with specific DNA sequences. Proteins are crosslinked in vivo with formaldehyde, and chromatin is then isolated and sheared. The protein of interest is then immunoprecipitated and the associated DNA sequences identified via PCR. Although this technique was originally designed to assay DNA binding proteins, it can also be used to monitor mRNA processing factors associated with transcription complexes.

摘要

染色质免疫沉淀(ChIP)技术已被用于确定DNA结合蛋白在何处以及在何种条件下与特定DNA序列结合。蛋白质在体内与甲醛交联,然后分离并剪切染色质。然后免疫沉淀感兴趣的蛋白质,并通过PCR鉴定相关的DNA序列。尽管该技术最初设计用于检测DNA结合蛋白,但它也可用于监测与转录复合物相关的mRNA加工因子。

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