| Metabolite Profiling of Mature Arabidopsis thaliana Seeds Using Gas Chromatography-Mass Spectrometry (GC-MS)
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Author:
Date:
2016-11-05
[Abstract] Metabolite profiling using gas chromatography-mass spectrometry (GC-MS) permits the annotation and quantification of a relatively wide variety of metabolites, covering a wide range of biochemical groups of metabolites. Lisec et al. (2006) established a method for GC-MS profiling in plants. Based on this protocol, we provide here a detailed GC-MS-based metabolite profiling protocol to identify compounds belonging to several biochemical groups in the primary metabolism of mature Arabidopsis thaliana seeds (Cohen et al., 2014). The protocol uses methoxyamine hydrochloride and N-methyl-N-trimethylsilyltriflouroacetamide (MSTFA) as derivatization reagents, as previous studies indicated these are the most appropriate compounds for profiling of plant ...
[摘要] 使用气相色谱 - 质谱（GC-MS）的代谢物分析允许注释和量化相对多种代谢物，涵盖广泛的代谢物的生物化学组。 Lisec等人。 （2006）建立了一种在植物中进行GC-MS分析的方法。基于该方案，我们在这里提供详细的基于GC-MS的代谢物分析方案以鉴定属于成熟拟南芥种子的主要代谢中的几个生物化学基团的化合物（Cohen等人。，2014）。该方案使用甲氧基胺盐酸盐和N，N-甲基-N，N-三甲基甲硅烷基三氟乙酰胺（MSTFA）作为衍生化试剂，如先前的研究所表明的，这些是用于分析植物代谢物的最合适的化合物。该方案相对快速，提供可重复的结果，并且可以用于简化许多其它类型的植物组织的代谢物，只有很少的修改。在这种情况下，开发种子可以作为研究代谢调节的优秀系统，因为在它们的开发期间，储备化合物的大规模合成在严格的转录调节下发生控制并且与时间上不同的代谢开关相关。
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| A Gas Chromatography-Mass Spectrometry-Based Two Stage Assay for Measurement of in vitro myo-Inositol 3-phosphate Synthase (INO1) Activity
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Author:
Date:
2015-03-05
[Abstract] This method describes an in vitro assay for measuring INO1 enzyme activity (the conversion of glucose 6-phosphate to myo-inositol 3-phosphate) in cell-free extracts. The method was first described for Plasmodium falciparum cells in MacRae et al. (2014) and consists of two parts: Part 1 describes the assay itself while part 2 describes analysis of the myo-inositol 3-phosphate product using gas chromatography-mass spectrometry (GC-MS).
[摘要] 该方法描述了用于测量无细胞提取物中INO1酶活性（葡萄糖-6-磷酸转化为肌醇-3-磷酸）的体外测定。 该方法首先在MacRae等人（2014）中描述了恶性疟原虫细胞，并由两部分组成：第1部分描述测定本身，而第2部分描述分析 使用气相色谱 - 质谱（GC-MS）测定肌醇3-磷酸产物。
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| Metabolite and Fatty Acid Analysis of Yeast Cells and Culture Supernatants
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Author:
Date:
2014-09-05
[Abstract] Metabolite and fatty acid analysis play important roles in evaluating the metabolic state of microorganisms. To examine the growth state and metabolism response of cells to environmental stress or genetic modification, intracellular and extracellular metabolites including fatty acids are usually analyzed to help understand the cellular biochemical changes in microorganisms. In this protocol, gas chromatography-mass spectrometry based analysis was employed to investigate the fatty acids and other metabolites in yeast cells.
[摘要] 代谢物和脂肪酸分析在评估微生物的代谢状态中起重要作用。 为了检查细胞对环境胁迫或遗传修饰的生长状态和代谢反应，通常分析细胞内和细胞外代谢物（包括脂肪酸）以帮助理解微生物中的细胞生物化学变化。 在本协议中，采用基于气相色谱 - 质谱法的分析来研究酵母细胞中的脂肪酸和其他代谢物。
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