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Cork-borer set

软木钻头套

Company: Sigma-Aldrich
Catalog#: Z165220
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Ubiquitin Proteasome Activity Measurement in Total Plant Extracts
Author:
Date:
2017-09-05
[Abstract]  The fine-tuned balance of protein level, conformation and location within the cell is vital for the dynamic changes required for a cell to respond to a given stimulus. This requires the regulated turnover of damaged or short-lived proteins through the ubiquitin proteasome system (UPS). Thus, the protease activity of the proteasome is adjusted to meet the current demands of protein degradation via the UPS within the cell. We describe the adaptation of an intramolecular quenched fluorescence assay utilizing substrate-mimic peptides for the measurement of proteasome activity in total plant extracts. The peptide substrates contain donor-quencher pairs that flank the scissile bond. Following cleavage, the increase in dequenched donor emission of the product is subsequently measured over time ... [摘要]  细胞内的蛋白质水平,构象和位置的微调平衡对于细胞对给定刺激作出反应所需的动态变化是至关重要的。 这需要通过泛素蛋白酶体系统(UPS)调节受损或短寿命蛋白质的周转。 因此,调节蛋白酶体的蛋白酶活性以满足目前通过细胞内的UPS的蛋白质降解的需求。 我们描述了使用底物 - 模拟肽进行分子内淬灭荧光测定来测定总植物提取物中蛋白酶体活性的适应性。 肽底物含有侧向于剪切键的供体 - 猝灭剂对。 切割后,随后随时测量产物的去质子供体发射的增加,并用于计算相对蛋白酶体活性。
【背景】泛素蛋白酶体系统(UPS)是真核细胞中主要的蛋白质降解机制,因此,UPS对许多细胞过程的调节至关重要,包括信号传导,细胞周期,囊泡运输和免疫。用于营业额的蛋白质通过泛素的共价连接标记,然后被26S蛋白酶体降解。 26S蛋白酶体由两个亚颗粒组成,即20S核心蛋白酶(CP),其分隔蛋白酶活性位点和19S调节颗粒,其将适当的底物识别并转移到CP腔中进行分解。调节蛋白酶体活性以维持蛋白酶抑制以响应内部和外部条件的波动。我们最近显示,UPS涉及植物免疫的几个方面,一系列植物和动物病原体颠覆了UPS来增强其毒力(Üstünet al。,2013;üstünet al。,2014;ÜstünandBörnke,2015 ;Üstünet ...

Infection of Nicotiana benthamiana Plants with Potato Virus X (PVX)
Author:
Date:
2016-12-20
[Abstract]  Potato Virus X (PVX) is the type member of Potexvirus genus, a group of plant viruses with a positive-strand RNA genome (~6.4 kb). PVX is able to establish compatible infections in Nicotiana benthamiana, a commonly used host in plant virology, leading to mild symptoms, such as chlorotic mosaic and mottling. PVX has been widely used as a viral vector for more than two decades (Chapman et al., 1992; Baulcombe et al., 1995; Aguilar et al., 2015). It provides a feasible means for the systemic expression in plants of heterologous proteins, such as avirulence factors, proteins with pharmacological properties, etc., (Hammond-Kosack et al., 1995; Gleba et al., 2014), and also as a tool to help decipher the function ... [摘要]  马铃薯病毒X(PVX)是Potexvirus属(一组具有正链RNA基因组(〜6.4kb)的植物病毒)的类型成员。 PVX能够在烟草烟草(Nicotiana benthamiana)中建立相容的感染,这是常用于植物病毒学的宿主,导致轻度症状,如褪色马赛克和斑驳。 PVX已经被广泛用作病毒载体二十多年(Chapman等人,1992; Baulcombe等人,1995; Aguilar等人。它为异源蛋白质植物中的全身表达提供了可行的手段,例如无毒性因子,具有药理学性质的蛋白质,等等(Hammond-Kosack等人, 1995; Gleba等人,2014),并且还作为通过病毒诱导的基因沉默(VACS)(Lacomme和Chapman,2008)帮助解释植物中基因功能的工具。摩擦(A)和农用过滤(B)的两种不同的方案,即允许PVX在N中的有效乘法和传播。本文详细描述本文。摩擦方法需要先前感染的液体,并且通过诱导对叶组织的机械损伤来实现感染,从而允许病毒颗粒穿透植物表面。农杆菌过滤需要先前修饰的农杆菌携带并将PVX序列递送到植物细胞中。将土壤杆菌生长直至饱和,并用注射器将其浸入植物组织中建立感染。这两种方法中的任何一种都可以成功应用,选择应主要基于材料和时间的可用性,但建议在使用嵌合病毒时使用农杆菌滤过。

背景 ...

Luminol-based Assay for Detection of Immunity Elicitor-induced Hydrogen Peroxide Production in Arabidopsis thaliana Leaves
Author:
Date:
2015-12-20
[Abstract]  In Arabidopsis thaliana, one of the very early immune-related responses induced after elicitor perception is the oxidative burst, i.e., reactive oxygen species (ROS) generation including superoxide anion and hydrogen peroxide (H2O2). ROS production plays different roles in a wide range of biotic and abiotic stress responses, including the closure of stomata and the regulation of cell expansion. In particular, elicitor-induced H2O2 is produced mainly by the membrane localized NAD(P)H oxidases RESPIRATORY BURST OXIDASE HOMOLOGUE D and F. In this protocol, we describe a simple and reproducible luminol/peroxidase-based assay to detect and evaluate immunity-related accumulation of H2O2 produced in Arabidopsis ... [摘要]  在拟南芥中,在诱导剂感知之后诱导的非常早期的免疫相关反应之一是氧化爆发,即包括超氧阴离子和过氧化氢的活性氧(ROS)产生(H 2 O 2 O 2)。 ROS产生在宽范围的生物和非生物应激反应中起不同的作用,包括气孔的闭合和细胞扩增的调节。特别地,诱导剂诱导的H 2 O 2 O 2主要由膜定位的NAD(P)H氧化酶呼吸道刺激氧化酶同源物D和F产生。在该方案中,我们描述了简单和可再现的基于鲁米诺/过氧化物酶的测定,以检测和评价在拟南芥叶盘中产生的H 2 O 2亚型的免疫相关累积(OGs),鞭毛蛋白(flg22)或延伸因子 - 热不稳定的(EF-Tu-elf18)处理。该方法基于检测在H 2 O 2 O 2存在下在辣根过氧化物酶(HRP) - 催化的鲁米诺分子氧化后产生的激发发光氨分子发出的发光, sub>。发光的水平以及持续时间与由引发的叶盘产生的H 2 O 2 O 2的量成比例。

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