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Glycerol

甘油

公司名称: Sigma-Aldrich
产品编号: G7757
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Expression, Purification and Crystallization of Recombinant Arabidopsis Monogalactosyldiacylglycerol Synthase (MGD1)
Author:
Date:
2016-12-20
[Abstract]  In plant cells, galactolipids are predominant, representing up to 50% of the lipid content in photosynthetic tissues. Galactolipid synthesis is initiated by MGDG synthases (MGDs), which use UDP-galactose as a donor sugar and diacylglycerol (DAG) as acceptor, to form monogalactosyldiacylglycerol (MGDG). This protocol is used to produce a recombinant form of Arabidopsis thaliana (A. thaliana) monogalactosyldiacylglycerol synthase 1 (MGD1) protein, in Escherichia coli (E. coli), using a two-step chromatographic purification procedure. The protein is easily ... [摘要]  在植物细胞中,半乳糖脂是主要的,代表高达光合组织脂质含量的50%。通过使用UDP-半乳糖作为供体糖和二酰基甘油(DAG)作为受体的MGDG合成酶(MGD)启动半乳糖脂合成以形成单糖基二酰基甘油(MGDG)。该方案用于在大肠杆菌中产生拟南芥(Arabidopsis thaliana)(拟南芥)单糖半乳糖二酰基甘油合酶1(MGD1)蛋白的重组形式(大肠杆菌),使用两步色谱纯化方法。蛋白质容易表达和纯化至数量,适用于生物化学和结构研究。还描述了MGD1的结晶。

背景 以前在e中表达植物MGD的尝试。大肠杆菌显示约99%的重组蛋白质积累在包涵体中(Miège等,1999)。开发了使用洗涤剂或体外包涵体折叠方案的细菌膜的溶解,并产生足够的纯和活性级分,足以监测酶的活性,但不进行其结构研究(Nishiyama et ...

Plant Tissue Trypan Blue Staining During Phytopathogen Infection
Author:
Date:
2016-12-20
[Abstract]  In this protocol plant tissue is stained with trypan blue dye allowing the researcher to visualize cell death. Specifically this method avoids the use of the carcinogen compound chloral hydrate, making this classical method of staining safer and faster than ever. The protocol is applied specifically to detect cell death on Arabidopsis leaves during the course of infection with necrotrophic fungus Botrytis cinerea. [摘要]  在该方案中,植物组织用台盼蓝染料染色,使研究人员可视化细胞死亡。具体来说,这种方法避免了使用致癌物水合氯醛,使得这种经典染色方法的染色比以往更加安全和快速。该方案专门用于在坏死性真菌Botrytis cinerea感染过程中检测拟南芥叶片上的细胞死亡。

【背景】 检测死植物组织的最常见的方法之一是台盼蓝染色(Keogh等人,1980)。该重氮染料也用于组织学和药物中,以通过允许细胞死亡的观察来测量组织存活力(Keogh等人,1980; Cooksey,2014)。涉及台盼蓝染色的大多数微观方法需要长时间的清除步骤,使用水合氯醛(CHL),一种目前使用的小型有机化合物,如实验动物中的致癌物,麻醉剂和止痛剂(Keogh等, 1980年; Lu和Greco,2006; Salmon等人,1995)。 CHL未经美国FDA批准或欧盟EMA在任何医疗指征( http://www.accessdata.fda.gov/ )。只有250mg或50mg的合唱水合物足以分别产生成人或儿科镇静剂,其毒性也在新生儿中测定( http://www.drugs.com ...

Secretion Assay in Shigella flexneri
Author:
Date:
2014-11-20
[Abstract]  Shigella flexneri (S. flexneri) is a Gram-negative bacterium that causes gastroenteritis and shigellosis in humans. In order to establish and maintain an infection, S. flexneri utilises a type three secretion system (T3SS) to deliver virulence factors called effector proteins into the cytoplasm of host cells, facilitating e.g. uptake into the host cell and escape from the endosome. Secretion through the T3SS is tightly regulated and is usually triggered by host-cell contact, but can also be artificially stimulated in vitro. In this assay, the dye ... [摘要]  志贺氏菌(Shigella flexneri)( S.flexneri )是一种革兰氏阴性细菌,其在人类中引起胃肠炎和志贺氏菌病。为了建立和维持感染, flexneri利用三型分泌系统(T3SS)将称为效应子蛋白的毒力因子递送到宿主细胞的细胞质中,促进例如摄取到宿主细胞中并从内体中逃逸。通过T3SS的分泌是严格调节的,通常由宿主细胞接触引发,但也可以在体外人工刺激。在该测定中,染料刚果红用于诱导T3的依赖于T3SS的分泌。 (Parsot等人,1995),通过用三氯乙酸沉淀从培养物上清液中浓缩分泌的蛋白质。本文提供的测定可以容易地适应于利用T3SS(例如鼠伤寒沙门氏菌)的其他细菌的分泌分析,当在37℃下生长时,其组成性分泌(Collazo等, (Heesemann等人,1995; Pegues等人,1995)或Yersinia的致病物种,其中可通过钙剥夺诱导分泌(Heesemann等人,/em,1986; Forsberg等人,1987)。

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