Development of an efficient chromatin immunoprecipitation method to investigate protein-DNA interaction in oleaginous castor bean seeds
Haque, Mohammad Enamul1,2; Han, Bing1,2; Wang, Bin1,3; Wang, Yue1; Liu, Aizhong1,3
2018-05-08
发表期刊PLOS ONE
ISSN1932-6203
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.
语种英语
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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
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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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