| Chromatin Immunoprecipitation (ChIP) to Assess Histone Marks in Auxin-treated Arabidopsis thaliana Inflorescence Tissue
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Author:
Date:
2020-12-05
[Abstract] Chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) or high-throughput sequencing (ChIP-seq) has become the gold standard for the identification of binding sites of DNA binding proteins and the localization of histone modification on a locus-specific or genome-wide scale, respectively. ChIP experiments can be divided into seven critical steps: (A) sample collection, (B) crosslinking of proteins to DNA, (C) nuclear extraction, (D) chromatin isolation and fragmentation by sonication, (E) immunoprecipitation of histone marks by appropriate antibodies, (F) DNA recovery, and (G) identification of precipitated protein-associated DNA by qPCR or high-throughput sequencing. Here, we describe a time-efficient protocol that can be used for ChIP-qPCR experiments to study the ...
[摘要] [摘要]染色质免疫沉淀与定量PCR（ChIP -qPCR）或高通量测序（ChIP-seq ）结合已成为鉴定DNA结合蛋白结合位点和在特定基因座上定位组蛋白修饰的金标准。或全基因组规模。ChIP实验可分为七个关键步骤：（A）样品收集，（B）蛋白质与DNA交联，（C）核提取，（D）染色质分离和f 超声处理的碎片化；（E）通过适当的抗体对组蛋白标记的免疫沉淀；（F）DNA的回收；（G）通过qPCR或高通量测序鉴定沉淀的蛋白质相关DNA。在这里，我们描述了一种可用于ChIP -qPCR实验的省时协议，以研究模型植物拟南芥幼花序中组蛋白修饰的定位。
[背景]真核基因组中的染色体中，其与组蛋白DNA结合形成染色质组织的。组蛋白与DNA之间的紧密相互作用阻碍了DNA与其他因素的可及性。因此，组蛋白相对于重要调控DNA序列的位置和组蛋白-DNA接触的强度可以隐藏或暴露提供另一层基因调控的基因。在染色质中，组蛋白和DNA均可被化学修饰（Zhou等，2010 ；Schübeler ，2015）。根据修饰的物理性质，染色质状态可以阻止或增强基础基因的转录（Kouzarides ，2007； Yang等，2014； Wu等，2015）。在植物中，染色质的表观遗传状态已被证明是响应发育或环境刺激的基因表达的关键决定因素（Yang等人，2014 ; Wu等人，2015 ; ...
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| Growth Assay and Detection of TRP and Indole Derivatives in Piriformospora indica Culture Supernatant by LC-MS/MS
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Author:
Date:
2013-06-20
[Abstract] The mutualistic root endophyte Piriformospora indica colonizes a wide range of plants and the colonization of root cells by this fungus is very often associated with beneficial effects to its host, such as growth promotion and increased biotic and abiotic stress tolerance. These traits may be based on general mechanisms and signaling pathways common to many different plant species. One such mechanism could be the recruitment of phytohormone pathways by P. indica. It is known, that many mutualistic microorganisms are able to synthesize and secrete phytohormones during the interaction with their host plants. This protocol has been successfully utilized to analyze tryptophan (TRP)-dependent biosynthesis of indole-3-acetic acid (IAA) and its indole derivatives by P. indica ...
[摘要] 共生根内生真菌（Pyriformospora indica）殖民广泛的植物，并且这种真菌对根细胞的定植通常与对其宿主的有益效果相关，例如生长促进和增加的生物和非生物胁迫耐受性 。 这些性状可以基于许多不同植物物种共有的一般机制和信号传导途径。 一种这样的机制可以是通过P募集植物激素途径。 indica 。 已知许多共生微生物能够在与其宿主植物相互作用期间合成和分泌植物激素。 该方案已经成功地用于通过P分析吲哚-3-乙酸（IAA）及其吲哚衍生物的色氨酸（TRP）依赖性生物合成。 蚜虫以及它们对这种真菌生长的影响（Hilbert等人，2012）。
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| Indole Derivative Feeding Test and Detection of TRP and Indole derivatives by Thin Layer Chromatography
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Author:
Date:
2013-06-20
[Abstract] The mutualistic root endophyte Piriformospora indica colonizes a wide range of plants and the colonization of root cells by this fungus is very often associated with beneficial effects to its host, such as growth promotion and increased biotic and abiotic stress tolerance. These traits could be based on general mechanisms and signaling pathways common to many different plant species. One such mechanism could be the recruitment of phytohormone pathways by P. indica. It is known, that many mutualistic microorganisms are able to synthesize and secrete phytohormones during the interaction with their host plants. This protocol has been successfully utilized to analyze tryptophan (TRP)-dependent biosynthesis of indole-3-acetic acid (IAA) and its indole derivatives by P. indica ...
[摘要] 共生根内生真菌（Pyriformospora indica）殖民广泛的植物，并且这种真菌对根细胞的定植通常与对其宿主的有益效果相关，例如生长促进和增加的生物和非生物胁迫耐受性 。 这些性状可以基于许多不同植物物种共有的一般机制和信号途径。 一种这样的机制可以是通过P募集植物激素途径。 indica 。 已知许多共生微生物能够在与其宿主植物相互作用期间合成和分泌植物激素。 该方案已经成功地用于通过P分析吲哚-3-乙酸（IAA）及其吲哚衍生物的色氨酸（TRP）依赖性生物合成。 疟疾（Hilbert et al 。，2012）。
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