Expression, Purification and Crystallization of Recombinant Arabidopsis Monogalactosyldiacylglycerol Synthase (MGD1)
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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 ...
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An Assay to Study Botrytis cinerea-infected Grapevine Leaves Primed with Pseudomonas fluorescens
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Author:
Date:
2016-10-05
[Abstract] Grapevine (Vitis vinifera L.) is susceptible to an array of diseases among them the grey mold caused by the necrotrophic fungus Botrytis cinerea that decreases grape productivity and quality. To ensure a satisfactory yield and harvest quality numerous chemical fungicides are required, but they have serious drawbacks. One alternative is the use of beneficial bacteria to improve plant health. Pseudomonas fluorescens has been shown to trigger a plant-mediated resistance response in aboveground plant tissues against fungal, oomycete, bacterial, and viral pathogens. ...
[摘要] 葡萄(葡萄(Vitis vinifera)L.)易受一系列疾病的影响,其中包括由坏死性真菌葡萄孢菌引起的灰霉病,其降低葡萄生产力和质量。为了确保令人满意的产量和收获质量,需要许多化学杀真菌剂,但它们具有严重的缺点。一个替代方案是使用有益细菌来改善植物健康。已经显示荧光假单胞菌在地上植物组织中触发植物介导的对真菌,卵菌,细菌和病毒病原体的抗性反应。触发的植物抗性通过启动状态利用植物免疫系统的机制,其提供植物在病原体感染后快速和强烈地激活防御反应的增强能力,导致较低的适合度成本。有益细菌的引发应答包括防御相关基因的诱导表达,细胞壁增强和病原体感染后次生代谢产物的产生。在该方案中,我们描述了根据Verhagen评价有益细菌荧光假单胞菌PTA-CT2对葡萄植物的引发状态及其对灰葡萄孢的抗性水平的实验设计等。 (2011)和Gruau等人。 (2015)。
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