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Author:
Date:
2016-08-20
[Abstract] VPS34 is the only class III phosphatidylinositol-3-kinase (PI3K) in mammalian cells and produces the vast majority of cellular phosphatidylinositol-3-phosphate [PI(3)P]. PI(3)P is a key signalling lipid that plays many membrane trafficking roles in processes such as endocytosis and autophagy. VPS34 is a key cellular regulator, loss of function can have catastrophic effects and is embryonic lethal (Zhou et al., 2011). The levels of cellular PI(3)P can be determined by fluorescent staining techniques and can be used to monitor effects upon VPS34 activity, however it is important to verify that any changes are mediated by VPS34, particularly as alternate pathways of PI(3)P production are possible such as via class II PI3Ks (Devereaux et al., 2013). Assaying VPS34 activity ...
[摘要] VPS34是哺乳动物细胞中唯一的III类磷脂酰肌醇-3-激酶（PI3K），并且产生绝大多数细胞磷脂酰肌醇-3-磷酸[PI（3）P]。 PI（3）P是一个关键的信号脂质，在诸如内吞作用和自噬的过程中起许多膜运输的作用。 VPS34是关键的细胞调节剂，功能丧失可具有灾难性作用并且是胚胎致死的（Zhou等人，2011）。 细胞PI（3）P的水平可以通过荧光染色技术确定，并且可以用于监测对VPS34活性的影响，然而重要的是验证任何变化由VPS34介导，特别是作为PI（3）的替代途径， P生产是可能的，例如通过II类PI3K（Devereaux等人，2013）。 直接在体外测定VPS34活性可以是描绘特定刺激的作用的关键阶段。
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Author:
Date:
2013-06-05
[Abstract] The primary aerial surfaces of all land plants are coated by a lipidic cuticle, which restricts non-stomatal water loss and protects the plant from pathogens, herbivores, and ultraviolet radiation. The cuticle is made up of two components: cutin, a polymer of hydroxy- and epoxy- long-chain fatty acid derivatives and glycerol, and cuticular waxes, which are derivatives of very-long-chain fatty acids. While chemical analysis of cutin can be a lengthy and technically challenging task, analysis of cuticular waxes is relatively simple, and can be routinely used to characterize different plant species, adaptations of a given species to environmental conditions, or mutant phenotypes. Here, we present a protocol tailored for the analysis of cuticular waxes on the surface of the model organism Arabidopsis ...
[摘要] 所有陆地植物的主要空间表面都涂有脂质角质层，限制非气孔水分流失，保护植物免受病原体，食草动物和紫外线辐射。角质层由两部分组成：角质，羟基和环氧长链脂肪酸衍生物的聚合物和甘油，以及作为非常长链脂肪酸衍生物的角质蜡。虽然角质层的化学分析可能是一个漫长而技术上具有挑战性的任务，但是角质层蜡的分析相对简单，可以常规用于表征不同植物物种，给定物种适应环境条件或突变体表型。在这里，我们提出了一种针对拟南芥模拟生物表面上的角质层蜡分析的方案。因为在植物的最外表面上发现角质蜡，蜡提取过程非常简单，样品处理可在不到一天的时间内完成。化学分析包括通过气相色谱与火焰离子化检测（GC / FID）联用对蜡单体进行定量，以及通过质谱法或单个蜡组分与已知标准物的保留时间的比较来鉴定蜡单体。
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