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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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