Tissue specificity and developmental pattern of amorpha-4,11-diene synthase (ADS) proved by ADS promoter-driven GUS expression in the heterologous plant, Arabidopsis thaliana
Tissue specificity and developmental pattern of amorpha-4,11-diene synthase (ADS) proved by ADS promoter-driven GUS expression in the heterologous plant, Arabidopsis thaliana
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DOI：
10.1055/s-2008-1034276
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发表时间：
2008-02-01
期刊：
影响因子：
2.7
通讯作者：
Kim, Soo-Un
中科院分区：
文献类型：
--
作者：
Kim, Soon-Hee;Chang, Yung-Jin;Kim, Soo-Un
青蒿的 Amorpha-4,11-diene 合酶 (ADS) 是一种倍半萜环化酶，在抗疟青蒿素的生物合成中催化法呢基二磷酸转化为 Amorpha-4,11-diene。为了探索调节 ADS 的组织特异性和发育分布的机制，使用 PCR 生成了完整的 ADS 启动子，并将其与 GUS 融合以引入拟南芥中。 ADSpro::GUS 融合转录本具有器官特异性，主要存在于幼叶绿色组织的花药和毛状体中。该结果与通过 RT-PCR 检查在青蒿中观察到的 ADS 转录模式一致。为了确定 ADS 的亚细胞定位，将 ADS 的开放阅读框 (ORF) 与绿色荧光蛋白 (smGFP) 基因融合并引入拟南芥原生质体中。 GFP 荧光仅位于细胞质中，表明 ADS 是一种细胞质定位蛋白。
Amorpha-4,11-diene synthase (ADS) of Artemisia annua L. is a sesquiterpene cyclase that catalyzes the conversion of farnesyl diphosphate into amorpha-4,11-diene in the biosynthesis of the antimalarial artemisinin. To explore the mechanisms regulating the tissue-specific and developmental distributions of ADS, a full ADS promoter was generated using PCR, and fused to GUS for introduction into Arabidopsis thaliana. ADSpro::GUS fusion transcripts were organ-specific, mainly present in the anthers and trichomes of the green tissues of the juvenile leaves. This result was consistent with the ADS transcription pattern observed in A. annua as examined by RT-PCR. To determine the subcellular localization of ADS, an open reading frame (ORF) of ADS was fused to the green fluorescent protein (smGFP) gene and introduced into the A. thaliana protoplasts. GFP fluorescence was located exclusively in the cytosol, an indication that ADS is a cytosol-localized protein.