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毛果巴豆中普伐他汀生物合成途径中编码1-脱氧-D-木酮糖-5-磷酸合酶和2-C-甲基-D-赤藓糖醇-4-磷酸合酶基因的转录谱分析

Transcription profiles analysis of genes encoding 1-deoxy-D-xylulose 5-phosphate synthase and 2C-methyl-D-erythritol 4-phosphate synthase in plaunotol biosynthesis from Croton stellatopilosus.

作者信息

Wungsintaweekul Juraithip, Sirisuntipong Tanawan, Kongduang Damrong, Losuphanporn Thanaisawan, Ounaroon Anan, Tansakul Pimpimon, De-Eknamkul Wanchai

机构信息

Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand.

出版信息

Biol Pharm Bull. 2008 May;31(5):852-6. doi: 10.1248/bpb.31.852.

Abstract

Genes encoding 1-deoxy-D-xylulose 5-phosphate synthase (DXS; EC 2.2.1.7) and 2C-methyl-D-erythritol 4-phosphate synthase (MEPS; EC 1.1.1.267), the first two enzymes in the deoxyxylulose phosphate (DXP) pathway, were cloned from young leaves of Croton stellatopilosus, and designated as 1-deoxy-D-xylulose 5-phosphate synthase (CSDXS) and 2C-methyl-D-erythritol 4-phosphate synthase (CSMEPS), respectively. Analysis of deduced amino acid sequences of the CSDXS and the CSMEPS confirmed their nucleotide sequences as they shared high identities to other known DXSs and MEPSs in higher plants. Physiological roles of the CSDXS and the CSMEPS were determined for the mRNA expressions in leaves, twigs and roots. Transcription profiles analyses of the CSDXS and the CSMEPS genes were investigated using semi-quantitative RT-PCR technique. Relative intensities of the CSDXS and the CSMEPS expressions to house-keeping gene (18S rRNA) were calculated. The results indicated that the levels of mRNAs expressions of the CSDXS and the CSMEPS were high in leaves and twigs. This evidence was in line with the high content of plaunotol, accumulated in leaves and twigs. Neither the CSDXS nor the CSMEPS were expressed in roots, where plaunotol was not detected. From this study, it can be concluded that plaunotol is biosynthesized in the chloroplastic tissue and regulated by the CSDXS and the CSMEPS.

摘要

编码1-脱氧-D-木酮糖-5-磷酸合酶(DXS;EC 2.2.1.7)和2-C-甲基-D-赤藓糖醇-4-磷酸合酶(MEPS;EC 1.1.1.267)的基因,即磷酸脱氧木酮糖(DXP)途径中的前两种酶,从毛果巴豆的幼叶中克隆得到,分别命名为1-脱氧-D-木酮糖-5-磷酸合酶(CSDXS)和2-C-甲基-D-赤藓糖醇-4-磷酸合酶(CSMEPS)。对CSDXS和CSMEPS推导的氨基酸序列分析证实了它们的核苷酸序列,因为它们与高等植物中其他已知的DXS和MEPS具有高度同源性。测定了CSDXS和CSMEPS在叶、嫩枝和根中的mRNA表达的生理作用。使用半定量RT-PCR技术研究了CSDXS和CSMEPS基因的转录谱分析。计算了CSDXS和CSMEPS表达相对于看家基因(18S rRNA)的相对强度。结果表明,CSDXS和CSMEPS的mRNA表达水平在叶和嫩枝中较高。这一证据与叶和嫩枝中积累的高含量稻瘟醇一致。在未检测到稻瘟醇的根中,CSDXS和CSMEPS均未表达。从这项研究可以得出结论,稻瘟醇在叶绿体组织中生物合成,并受CSDXS和CSMEPS调控。

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