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公司名称: Merck
产品编号: G8270
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Imaging Microtubules in vitro at High Resolution while Preserving their Structure
Author:
Date:
2021-04-05
[Abstract]  

Microtubules (MT) are the most rigid component of the cytoskeleton. Nevertheless, they often appear highly curved in the cellular context and the mechanisms governing their overall shape are poorly understood. Currently, in vitro microtubule analysis relies primarily on electron microscopy for its high resolution and Total Internal Reflection Fluorescence (TIRF) microscopy for its ability to image live fluorescently-labelled microtubules and associated proteins. For three-dimensional analyses of microtubules with micrometer curvatures, we have developed an assay in which MTs are polymerized

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[摘要]  [摘要]微管(MT)是细胞骨架中最刚性的组成部分。然而,它们在细胞环境中经常显得高度弯曲,并且控制它们整体形状的机理了解甚少。当前,体外微管分析主要依靠电子显微镜进行高分辨率分析,而全内反射荧光(TIRF )显微镜则可以对活的荧光标记的微管和相关蛋白进行成像。为了对具有微米曲率的微管进行三维分析,我们开发了一种在体外聚合MT的检测方法 用类似于常规TIRF显微镜操作规程的方式将MT种子的MT粘附到载玻片上。除去游离的荧光分子,并通过灌注固定MTs。然后可以使用带有Airyscan模块的共聚焦显微镜观察MT,以获得更高的分辨率。该协议允许对保留其原始三维形状并与高分辨率免疫荧光检测兼容的微管进行成像。

[背景]微管(MT)是通过异源二聚体的组合制成的聚合物α和β微管蛋白,并且是细胞骨架的主要成分。他们参与了细胞功能的基本机制,如有丝分裂,细胞内转运,胞质分裂和细胞形态的维持(Akhmanova和Steinmetz,2015)。尽管MT本身具有很高的刚性,但它们通常会在细胞中弯曲并产生一些蛋白,从而弯曲微管(Brangwynne等人,2006; Bechstedt等人,2014; Leung等人,2020; Cuveillier等人,2020 ...

Chromatographic Analysis for Targeted Metabolomics of Antioxidant and Flavor-Related Metabolites in Tomato
Author:
Date:
2021-03-05
[Abstract]  

Targeted metabolomics is a useful approach to evaluate crop breeding studies. Antioxidant and flavor-related traits are of increasing interest and are considered quality traits in tomato breeding. The present study presents chromatographic methods to study antioxidants (carotenoids, vitamin C, vitamin E, phenolic compounds, and glutathione) and flavor-related characters (sugars and organic acids) in tomato. Two different extraction methods (for polar and apolar entities) were applied to isolate the targeted compounds. The extraction methods developed in this work were time and cost-effective

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[摘要]  [摘要]靶向代谢麦克风是评估作物育种研究的有用方法。抗氧化剂和与风味有关的性状受到越来越多的关注,被认为是番茄育种中的品质性状。本研究提供了色谱方法来研究番茄中的抗氧化剂(类胡萝卜素,维生素C,维生素E,酚类化合物和谷胱甘肽)和与风味相关的特征(糖和有机酸)。应用了两种不同的提取方法(针对极性和非极性实体)来分离目标化合物。由于不需要进一步纯化,因此这项工作中开发的提取方法既省时又经济。使用RP C18色谱柱,通过RP-C18色谱柱在适当波长下对每种化合物进行类胡萝卜素,维生素C,谷胱甘肽和酚酸的分析。用RP C18和NH2色谱柱通过HPLC分析维生素E和糖,并分别通过FLD和RI检测器进行检测。此外,MSTFA衍生化后,使用Rtx 5DA色谱柱使用GC-FID分析有机酸。结果,开发了定量重要植物代谢物的灵敏分析方法,并在本文中进行了描述。这些方法不仅适用于番茄,但都还表征其他物种的香味相关的和抗氧化的化合物。因此,这些协议可用于指导作物育种的选择。

[背景]代谢组学是其受到关注的其潜在的援助作物育种研究施加生化途径。在番茄育种中,由于消费者对提高作物品质的偏好,抗氧化剂和与风味有关的代谢产物都受到了越来越多的关注。人类无法自己合成抗氧化剂分子; ...

EmPC-seq: Accurate RNA-sequencing and Bioinformatics Platform to Map RNA Polymerases and Remove Background Error
Author:
Date:
2021-02-20
[Abstract]  

Transcription errors can substantially affect metabolic processes in organisms by altering the epigenome and causing misincorporations in mRNA, which is translated into aberrant mutant proteins. Moreover, within eukaryotic genomes there are specific Transcription Error-Enriched genomic Loci (TEELs) which are transcribed by RNA polymerases with significantly higher error rates and hypothesized to have implications in cancer, aging, and diseases such as Down syndrome and Alzheimer’s. Therefore, research into transcription errors is of growing importance within the field of genetics.

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[摘要]  [摘要]转录错误可通过改变表观基因组并引起mRNA的错误整合而严重影响生物体内的代谢过程,从而将其翻译为异常的突变蛋白。此外,真核基因组内有特定转录错误富集的基因组基因座(TEELs),它们由RNA聚合酶与显著更高的错误率转录并推测为具有影响在癌症,老化和疾病例如唐氏综合征和阿尔茨海默'秒。因此,在遗传学领域对转录错误的研究越来越重要。尽管如此,方法上的障碍限制了准确识别转录错误的进展。Pro-Seq和NET-Seq可以沿基因组纯化新生RNA并绘制RNA聚合酶,但不能用于鉴定转录突变。在这里,我们本背景误差模型耦合的精密核圆形测序上运行(EMPC -SEQ),一种方法COMBIN荷兰国际集团测定和圆形测序核上运行与背景误差模型精确地检测新生转录错误和有效地辨别TEELs基因组中。

[背景]核糖核苷酸错掺导致的转录错误在所有活生物体中无处不在(Carey,2015)。假设每个信使RNA(mRNA)可以翻译2-4千次(Schwanhausser et al。,2011),并且许多特殊RNA在给定时间每个细胞仅表达一次(Islam et al。,2011; Pelechano et al。,2011)。,2010),即使是关键残基的单个转录错误也会使特定蛋白质的表达产生很大差异。另外,转录错误可加速蛋白质聚集,导致人类中与年龄有关的疾病(van ...

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