| Extraction and Analysis of Carotenoids from Escherichia coli in Color Complementation Assays
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Author:
Date:
2017-03-20
[Abstract] A common method to investigate the function of genes putatively involved in carotenoid biosynthesis is the so called color complementation assay in Escherichia coli (see, e.g., Cunningham and Gantt, 2007). In this assay, the gene under investigation is expressed in E. coli strains genetically engineered to synthesize potential carotenoid substrates, followed by analysis of the pigment changes in the carotenogenic bacteria via high-performance liquid chromatography (HPLC). Two crucial steps in this method are (i) the quantitative extraction of the carotenoids out of E. coli and (ii) the reproducible and complete separation of the pigments by HPLC.
Here, we present a protocol for the extraction and analysis of carotenoids with a broad range of ...
[摘要] 调查涉及类胡萝卜素生物合成的基因的功能的常见方法是在大肠杆菌中所谓的颜色互补测定（参见，例如，Cunningham和Gantt，2007） 。在该测定中，所研究的基因在E中表达。基因工程改造以合成潜在的类胡萝卜素底物，然后通过高效液相色谱（HPLC）分析色素变性细菌。该方法的两个关键步骤是（i）从E中定量提取类胡萝卜素。大肠杆菌和（ii）通过HPLC重现和完全分离颜料。
＆nbsp;在这里，我们提出了一种从胡萝卜素E提取和分析具有广泛极性的类胡萝卜素的方案。大肠杆菌。用于提取的溶剂混合物在溶液中保持亲脂性胡萝卜素和更极性的叶黄素，并且与随后的HPLC分析的洗脱液梯度相容。所用的C30柱特别适于分离类胡萝卜素的各种顺式异构体，而且用于分离立体异构体如α-和β-胡萝卜素或叶黄素和玉米黄质。
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| Phenol-based Total Protein Extraction from Lily Plant Tissues
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Author:
Date:
2015-02-20
[Abstract] The phenol-based total protein extraction method is unique in that water-soluble components such as polyphenolic compounds and nucleic acids can be easily removed. Thus, total protein is free from contaminants and allows for high quality two–dimensional gel electrophoresis. The phenol-based extraction of total protein was used in various lily organs and may likely apply to other plants whose content of polyphenolics is high (Note 1). An additional advantage of this extraction method is that nucleic acids can be easily removed and thus, avoid adverse effects of nucleic acids on protein resolution in the gel. This method is modified from that of Hurkman and Tanaka (1986).
[摘要] 酚类总蛋白提取方法的独特之处在于可以容易地除去多酚化合物和核酸等水溶性成分。 因此，总蛋白不含污染物，并允许高质量的二维凝胶电泳。 总蛋白的苯酚提取用于各种百合器官，可能适用于多酚含量高的其他植物（注1）。 该提取方法的另一个优点是，可以容易地除去核酸，从而避免核酸对凝胶中蛋白质分辨的不利影响。 这种方法从Hurkman和Tanaka（1986）的修改。
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| Extraction of Total Proteins from Rice Plant
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Author:
Date:
2014-11-05
[Abstract] This protocol provides an efficient method for preparation of high-quality proteins from rice leaves and grains. The method involves phenol extraction to separate proteins from the non-protein components such as polysaccharides, lipids and phenolic compounds that are commonly enriched in plant tissues. Following isolation, proteins are precipitated with ammonium acetate/methanol and then solubilized for proteomic analysis. As the protocol is simple, universal, and most importantly compatible with silver staining, it has been applied to our routine protein extraction from rice and many other plant tissues and it even works fine in animal tissues for the requirement of electrophoretic separation.
[摘要] 这个协议提供了一种从稻叶和谷物制备高质量蛋白质的有效方法。 该方法包括酚提取以从通常富含植物组织的非蛋白质组分如多糖，脂质和酚类化合物中分离蛋白质。 分离后，用乙酸铵/甲醇沉淀蛋白质，然后溶解用于蛋白质组分析。 由于协议简单，通用，最重要的是与银染色兼容，它已经应用于我们从水稻和许多其他植物组织的常规蛋白质提取，甚至在动物组织中对于电泳分离的要求很好。
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