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首页> 外文期刊>Analytical Biochemistry: An International Journal of Analytical and Preparative Methods >Assay for abscisic acid 8 '-hydroxylase activity of cloned plant cytochrome P450 oxidases in Saccharomyces cerevisiae
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Assay for abscisic acid 8 '-hydroxylase activity of cloned plant cytochrome P450 oxidases in Saccharomyces cerevisiae

机译:测定用于克隆植物的脱落酸8' - 羟基杂化酶活性酿酒酵母中克隆植物细胞色素P450氧化酶

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摘要

The plant hormone abscisic acid (ABA) regulates many processes, including response to drought, seed dormancy and abscission of leaves and fruits. For maintenance of ABA homeostasis, catabolism of ABA by 8'-hydroxylation and subsequent cyclisation to phaseic acid (PA) is crucial. However, detection of ABA 8'-hydroxylation activity is tedious. We present a simple and rapid method for detection of ABA 8'-hydroxylase activity by cloning cDNAs of interest and expressing the respective protein in yeast. Upon addition of ABA, PA is formed and subsequently quantified in the yeast cell culture supernatant by heart cutting 2D-HPLC or GC-MS.
机译:植物激素脱落酸(ABA)调节许多方法,包括对干旱,种子休眠和叶片和水果脱落的反应。 为了维持ABA稳态,ABA的分解代谢8'-羟基化和随后的分代酸(PA)的溶液至关重要。 然而,检测ABA 8'-羟基化活性乏味。 通过克隆感兴趣的CDNA和在酵母中表达各自的蛋白质,介绍了一种简单且快速的方法,用于通过克隆感兴趣的CDNA检测ABA 8'-羟化酶活性。 在添加ABA时,通过心脏切割2D-HPLC或GC-MS在酵母细胞培养上清液中形成并随后在酵母细胞培养上清液中定量PA。

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