| Purification of Starch Granules from Arabidopsis Leaves and Determination of Granule-Bound Starch Synthase Activity
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Author:
Date:
2014-12-05
[Abstract] Starch constitutes the most important carbon reserve in plants and is composed of branched amylopectin and linear amylose. The latter is synthesized exclusively by the Granule-Bound Starch Synthase (GBSS, EC 2.4.1.21). Here we report a readily reproducible, specific and highly sensitive protocol, which includes the isolation of intact starch granules from Arabidopsis thaliana leaves and the subsequent determination of GBSS activity. We have applied this method to study GBSS activity in diurnal cycles in vegetative growth and during the photoperiodic transition to flowering in Arabidopsis (Tenorio et al., 2003; Ortiz-Marchena et al., 2014).
[摘要] 淀粉构成植物中最重要的碳储量，由支链支链淀粉和线性直链淀粉组成。 后者仅由颗粒结合的淀粉合酶（GBSS，EC 2.4.1.21）合成。 在这里我们报告一个容易重现，具体和高度敏感的协议，其中包括完整的淀粉颗粒从拟南芥叶中分离和随后的GBSS活动的确定。 我们已经应用这种方法来研究营养生长的日变化周期中和在拟南芥中的光周期过渡到开花期间的GBSS活性（Tenorio等人，2003; Ortiz-Marchena& em> et al。，2014）。
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| Metabolic Labeling of Yeast Sphingolipids with Radioactive D-erythro-[4,5-3H]dihydrosphingosine
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Author:
Date:
2013-08-20
[Abstract] Yeast sphingolipids can be metabolically labeled with exogenously added radioactive precursors. Here we describe a method to label ceramides and complex sphingolipids of Saccharomyces cerevisiae with a radioactive ceramide precursor, D-erythro-[4, 5-3H]dihydrosphingosine. This protocol is used to study the biosynthesis, transport and metabolism of sphingolipids in yeast.
[摘要] 酵母鞘脂可以用外源添加的放射性前体代谢标记。 在这里我们描述了用放射性神经酰胺前体D-赤式 - [4,5] 3 H]二氢鞘氨醇标记神经酰胺和酿酒酵母的复杂神经鞘脂的方法。 该方案用于研究酵母中鞘脂的生物合成，转运和代谢。
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