Protocol: methodology for chromatin immunoprecipitation (ChIP) in Chlamydomonas reinhardtii

被引：22

作者：

Strenkert, Daniela ^{[1
]}

Schmollinger, Stefan ^{[1
]}

Schroda, Michael ^{[1
]}

机构：

[1] Max Planck Inst Mol Pflanzenphysiol, D-14476 Potsdam, Germany

来源：

PLANT METHODS | 2011年 / 7卷

关键词：

Chromatin immunoprecipitation; Chlamydomonas reinhardtii; heat shock; formaldehyde crosslinking; real-time PCR; nucleosome occupancy; histone modification; HSP70A; RBCS2; CYC6; CARBON-CONCENTRATING MECHANISM; PROTEIN DNA INTERACTIONS; HEAT-SHOCK; GENE; FORMALDEHYDE; ACETYLATION; REGULATOR; REGIONS; CORE;

D O I：

10.1186/1746-4811-7-35

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We report on a detailed chromatin immunoprecipitation (ChIP) protocol for the unicellular green alga Chlamydomonas reinhardtii. The protocol is suitable for the analysis of nucleosome occupancy, histone modifications and transcription factor binding sites at the level of mononucleosomes for targeted and genome-wide studies. We describe the optimization of conditions for crosslinking, chromatin fragmentation and antibody titer determination and provide recommendations and an example for the normalization of ChIP results as determined by real-time PCR.

引用

页数：9

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