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3',5'-Dimethoxy-4'-hydroxyacetophenone

3'',5''-二甲氧基-4''-羟基苯乙酮

Company: Sigma-Aldrich
Catalog#: D134406
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Histone Deubiquitination Assay in Nicotiana benthamiana
Author:
Date:
2018-03-05
[Abstract]  Histone modifications are a group of post-translational modifications on histones which can alter chromatin structure and affect gene expression. Histone ubiquitination is a histone modification found in particular on histone H2A and H2B. Histone ubiquitination can be reversed by ubiquitin-specific proteases (UBP). Here, we describe an in vivo assay for histone deubiquitination activity. After infiltrating UBP12 into Nicotiana benthamiana leaves, H2Aub was visualized by immunocytochemistry. Nicotiana benthamiana leaves, which show high agro infiltration efficiency, were used for transient UBP12 expression for a labor- and time-saving protocol. Reduced H2Aub levels indicated histone deubiquitination activity of UBP12. The clear visualization of nuclei of N. ... [摘要]  组蛋白修饰是一组组蛋白翻译后修饰,可以改变染色质结构并影响基因表达。组蛋白泛素化是组蛋白H2A和H2B特异性发现的组蛋白修饰。泛素特异性蛋白酶(UBP)可以逆转组蛋白泛素化。在这里,我们描述了组蛋白去泛素化活性的体内试验。在将UBP12渗入烟草叶片中后,通过免疫细胞化学观察H2Aub。表现出高的农业渗透效率的本氏烟草叶用于瞬时UBP12表达,用于节省劳力和时间的方案。 H2Aub水平降低表明UBP12的组蛋白去泛素化活性。 N的核的清晰可视化。本生叶使得该方法能够通过使用特异性抗体容易地测量体内组蛋白修饰的水平,从而提供强大的蛋白质功能线索。因此,该协议是组蛋白去泛素化活性的体外试验的有力补充。

【背景】组蛋白修饰在调节染色质结构和基因表达中发挥重要作用。 研究最深入的组蛋白修饰包括甲基化,乙酰化,磷酸化,泛素化和sumoylation。 然而,引入或去除特定组蛋白修饰的酶并不总是已知的。 强大的体外试验可以确定组蛋白修饰酶的催化潜能,但是体内试验方法对于确认体外试验的特异性反映抗体的特异性是必要的。 体内活动。 在这里,我们描述了一个灵活的协议来测试植物组织中组蛋白修饰酶的活性。本塞姆氏。 尽管我们使用该方案来测试遍在蛋白特异性蛋白酶(UBP)对泛素化H2A的活性,但它也可以容易地用于其他特异性抗体可用的组蛋白修饰。

Trimolecular Fluorescence Complementation (TriFC) Assay for Direct Visualization of RNA-Protein Interaction in planta
Author:
Date:
2017-10-20
[Abstract]  RNA-Protein interactions play important roles in various eukaryotic biological processes. Molecular imaging of subcellular localization of RNA/protein complexes in plants is critical for understanding these interactions. However, methods to image RNA-Protein interactions in living plants have not yet been developed until now. Recently, we have developed a trimolecular fluorescence complementation (TriFC) system for in vivo visualization of RNA-Protein interaction by transient expression in tobacco leaves. In this method, we combined conventional bimolecular fluorescence complementation (BiFC) system with MS2 system (phage MS2 coat protein [MCP] and its binding RNA sequence [MS2 sequence]) (Schonberger et al., 2012). Target RNA is tagged with 6xMS2 and MCP and RNA binding ... [摘要]  RNA-蛋白质相互作用在各种真核生物过程中起重要作用。 RNA /蛋白质复合物在植物中亚细胞定位的分子成像对于理解这些相互作用至关重要。然而,到目前为止,尚未开发在活植物中形成RNA-蛋白质相互作用的方法。最近,我们开发了一种三分子荧光互补(TriFC)系统,用于在烟草叶中瞬时表达的RNA-蛋白质相互作用的体内可视化。在这种方法中,我们将传统的双分子荧光互补(BiFC)系统与MS2系统(噬菌体MS2外壳蛋白[MCP]及其结合RNA序列[MS2序列])(Schonberger等人,2012)相结合, 。目标RNA用6xMS2标记,MCP和RNA结合蛋白与YFP片段融合。编码这种融合RNA和蛋白质的DNA构建体用土壤杆菌悬浮液渗入烟草叶中。通过共焦显微镜观察体内的RNA-蛋白质相互作用
【背景】近来,多种类型的长非编码RNA(lncRNA)已经被鉴定并显示出在转录调节和染色质修饰中起重要作用(St Laurent等人,2015)。到目前为止,lncRNA介导的功能的大多数分子机制与RNA-蛋白质相互作用密切相关(St ...

Ensifer-mediated Arabidopsis thaliana Root Transformation (E-ART): A Protocol to Analyse the Factors that Support Ensifer-mediated Transformation (EMT) of Plant Cells
Author:
Date:
2017-10-05
[Abstract]  Ensifer adhaerens OV14, a soil borne alpha-proteobacteria of the Rhizobiaceae family, fortifies the novel plant transformation technology platform termed ‘Ensifer-mediated transformation’ (EMT). EMT can stably transform both monocot and dicot species, and the host range of EMT is continuously expanding across a diverse range of crop species. In this protocol, we adapted a previously published account that describes the use of Arabidopsis thaliana roots to investigate the interaction of A. thaliana and Agrobacterium tumefaciens. In our laboratory, we routinely use A. thaliana root explants to examine the factors that enhance the utility of EMT. In addition, the E-ART protocol can be used to study the transcriptional response of E. ... [摘要]  OV14;土壤传播的根瘤菌科的α-变形细菌强化了新型植物转化技术平台,称为“插入式”介导的转化(EMT)。 EMT可以稳定地转化单子叶植物和双子叶植物,并且EMT的宿主范围在不同范围的作物种类上不断扩大。在这个协议中,我们调整了一个以前发布的帐户,描述了使用拟南芥根系来研究 A的相互作用。 thaliana 和根癌土壤杆菌。在我们的实验室,我们通常使用 A。 thaliana 根外植体,以检查增强EMT效用的因素。此外,E-ART协议可用于研究E的转录反应。接种外植体组织后的寄主植物,宿主植物,不同的引物菌株/突变体的可变性以及测试A的易感性。作为破译支持EMT的机制的手段。【背景】推进“Ensifer”介导的转化(EMT)技术以成功地转化双子叶菊,即拟南芥,马铃薯Solanum tuberosum ,Nicotiana tabacum ,Manihot esculenta ,欧洲油菜和单子叶植物;之前曾报道过(Wendt等人,2012; Zuniga-Soto等人),2015; Chavarriaga-Aguirre et al。,2016; Rathore等人,2016)。另外,E的基因组分析。 (2014)发现,该细菌具有7.7Mb的基因组,其包含两条环状染色体(3.96Mb和2.01Mb)和两条质粒(1.61Mb和125Kb) )。 ...

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