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Effect of an Introduced Phytoene Synthase Gene Expression on Carotenoid Biosynthesis in the Marine Diatom Phaeodactylum tricornutum

机译:引入的植物保护区合酶基因表达对海洋硅藻Phaeodactylum tricornutum中类胡萝卜素生物合成的影响。

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摘要

Carotenoids exert beneficial effects on human health through their excellent antioxidant activity. To increase carotenoid productivity in the marine Pennales Phaeodactylum tricornutum, we genetically engineered the phytoene synthase gene (psy) to improve expression because RNA-sequencing analysis has suggested that the expression level of psy is lower than other enzyme-encoding genes that are involved in the carotenoid biosynthetic pathway. We isolated psy from P. tricornutum, and this gene was fused with the enhanced green fluorescent protein gene to detect psy expression. After transformation using the microparticle bombardment technique, we obtained several P. tricornutum transformants and confirmed psy expression in their plastids. We investigated the amounts of PSY mRNA and carotenoids, such as fucoxanthin and β-carotene, at different growth phases. The introduction of psy increased the fucoxanthin content of a transformants by approximately 1.45-fold relative to the levels in the wild-type diatom. However, some transformants failed to show a significant increase in the carotenoid content relative to that of the wild-type diatom. We also found that the amount of PSY mRNA at log phase might contribute to the increase in carotenoids in the transformants at stationary phase.
机译:类胡萝卜素具有出色的抗氧化活性,对人体健康具有有益的作用。为了提高海洋Pennales Phaeodactylum tricornutum中类胡萝卜素的生产力,我们对八氢番茄红素合酶基因(psy)进行了基因工程改造,以提高表达水平,因为RNA测序分析表明psy的表达水平低于与psy参与的其他酶编码基因的表达水平。类胡萝卜素的生物合成途径。我们从角果疟原虫中分离了psy,并将该基因与增强的绿色荧光蛋白基因融合以检测psy表达。使用微粒轰击技术进行转化后,我们获得了几种角果疟原虫转化体,并确认了其质体中的psy表达。我们调查了不同生长阶段的PSY mRNA和类胡萝卜素(例如岩藻黄质和β-胡萝卜素)的量。与野生型硅藻中的水平相比,psy的引入使转化体中岩藻黄质的含量增加了约1.45倍。然而,相对于野生型硅藻,一些转化体未能显示出类胡萝卜素含量的显着增加。我们还发现在对数期PSY mRNA的量可能有助于平稳期转化子中类胡萝卜素的增加。

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