Development of an efficient chromatin immunoprecipitation method to investigate protein-DNA interaction in oleaginous castor bean seeds

doi:10.1371/journal.pone.0197126

	Development of an efficient chromatin immunoprecipitation method to investigate protein-DNA interaction in oleaginous castor bean seeds
	Haque, Mohammad Enamul 1,2; Han, Bing 1,2; Wang, Bin1,3 ; Wang, Yue 1; Liu, Aizhong1,3
	2018-05-08
发表期刊	PLOS ONE
ISSN	1932-6203
卷号	13 期号:5 页码:197126
摘要	Chromatin immunoprecipitation (ChIP) is usually a reliable technique to find the binding sites of a transcription factor. In the current study, we developed a suitable ChIP method using developing castor bean seeds. A castor bean seed with large and persistent endosperm contains high amounts of storage lipids (ca. 50-60%) and is often considered as a model material to studying seed biology. In oleaginous seeds, due to the rich oils which could seriously affect immunoprecipitation and DNA isolation, it is often difficult to carry out a successful ChIP experiment. Thus, the development of an efficient ChIP method for oleaginous seeds is required. In this study, we modified different steps, including tissue preparation for cross-linking, chromatin washing, sonication and immunoprecipitation of other existing methods. As exemplified by the targeted gene identification of a master regulator WRI1, which regulates fatty acid biosynthesis, we found that the improved ChIP method worked well. We analyzed percentage input and fold changes of the ChIPed DNA. We also made successful ChIP-seq libraries using this method. This method provides a technical support not only for use on castor bean seeds; it might be used equally to analyze protein-DNA interaction in vivo in other oleaginous seeds.
DOI	10.1371/journal.pone.0197126
语种	英语
引用统计
文献类型	期刊论文
条目标识符	http://ir.kib.ac.cn/handle/151853/61246
专题	资源植物与生物技术所级重点实验室
作者单位	1.Chinese Acad Sci, Kunming Inst Bot, Key Lab Econ Plants & Biotechnol, Yunnan Key Lab Wild Plant Resources, Kunming, Yunnan, Peoples R China 2.Univ Chinese Acad Sci, Beijing, Peoples R China 3.Southwest Forestry Univ, Key Lab Forest Resources Conservat & Utilizat Sou, Minist Educ, Kunming, Yunnan, Peoples R China
推荐引用方式 GB/T 7714	Haque, Mohammad Enamul,Han, Bing,Wang, Bin,et al. Development of an efficient chromatin immunoprecipitation method to investigate protein-DNA interaction in oleaginous castor bean seeds[J]. PLOS ONE,2018,13(5):197126.
APA	Haque, Mohammad Enamul,Han, Bing,Wang, Bin,Wang, Yue,&Liu, Aizhong.(2018).Development of an efficient chromatin immunoprecipitation method to investigate protein-DNA interaction in oleaginous castor bean seeds.PLOS ONE,13(5),197126.
MLA	Haque, Mohammad Enamul,et al."Development of an efficient chromatin immunoprecipitation method to investigate protein-DNA interaction in oleaginous castor bean seeds".PLOS ONE 13.5(2018):197126.

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