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A novel method for screening a vertebrate transcriptome for genes involved in carotenoid binding and metabolism

机译:一种筛选脊椎动物转录组中涉及类胡萝卜素结合和代谢的基因的新方法

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摘要

Carotenoid-based colour signals are widespread in the animal kingdom and common textbook examples of sexually selected traits. Carotenoid pigments must be obtained through the diet as all animals lack the enzymatic machinery necessary to synthesize them from scratch. Once ingested, carotenoids are metabolized, stored, transported and deposited, and some or all of these processes may be limiting for signal production and thus subjected to social or sexual selection on phenotypic coloration. Very little is known about which genes and physiological pathways are involved in carotenoid pigmentation which is unfortunate, as genetic information would allow us to investigate the biochemical consequences of sexual selection. In this study, we present a transcriptome-screening technique and apply it to a carotenoid-signalling bird species, the southern red bishop Euplectes orix, to uncover the gene(s) responsible for the conversion of dietary β-carotene (orange) to canthaxanthin (bright red). The transcriptome, extracted from the liver of a male entering his breeding moult, is expressed within bacterial cells genetically modified to synthesize beta-carotene. Effects of expressed E. orix proteins on the structure or amount of β-carotene are initially detected by eye (based on colour change) and subsequently confirmed by high-performance liquid chromatography. Here, we demonstrate the validity of the technique and provide a list of candidate genes involved in the carotenoid pigmentation pathway. We believe that this method could be applied to other species and tissues and that this may help researchers uncover the genetic basis of carotenoid coloration in vertebrates.
机译:基于类胡萝卜素的颜色信号广泛存在于动物界和有关性选择特征的普通教科书示例中。类胡萝卜素色素必须通过饮食获得,因为所有动物都缺乏从头合成色素的必要酶促机制。摄入后,类胡萝卜素被代谢,储存,运输和沉积,其中某些或全部过程可能会限制信号的产生,因此在表型着色时会受到社会或性别的选择。关于哪些基因和生理途径参与类胡萝卜素色素沉着是非常不幸的,这是不幸的,因为遗传信息将使我们能够调查性选择的生化后果。在这项研究中,我们提出了一种转录组筛选技术,并将其应用于类胡萝卜素信号鸟,南部的红色主教Euplectes orix,以发现负责将饮食中β-胡萝卜素(橙色)转化为角黄素的基因。 (亮红)。从进入其繁殖期的雄性肝脏中提取的转录组在经过基因修饰以合成β-胡萝卜素的细菌细胞中表达。首先用肉眼(基于颜色变化)检测表达的大肠杆菌蛋白对β-胡萝卜素的结构或数量的影响,然后通过高效液相色谱法确认。在这里,我们证明了该技术的有效性,并提供了涉及类胡萝卜素色素沉着途径的候选基因的列表。我们认为该方法可以应用于其他物种和组织,这可能有助于研究人员发现脊椎动物类胡萝卜素着色的遗传基础。

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