作 者 :张萌,苏平,刘雨佳,童宇茹,赵瑜君,高伟,王秀娟,黄璐琦
期 刊 :中国中药杂志 2015年 40卷 6期 页码:1066.
关键词:雷公藤 ;牻牛儿基牻牛儿基焦磷酸合酶 ;克隆 ;生物信息学分析 ;
Keywords:Tripterygium wilfordii , GGPPS , clone , bioinformatics analysis ,
摘 要 :该研究利用cDNA末端快速扩增(RACE)技术从雷公藤悬浮细胞中克隆得到一条GGPPS 全长cDNA(GeneBank:KM978333),并对其进行多重序列比对、蛋白结构预测及构建系统进化树等生物信息学分析。所克隆的TwGGPPS cDNA全长1 857 bp,编码含514个氨基酸的蛋白序列,相对分子质量为57.13 kDa,等电点为7.85,具有5个保守域和2个功能域,亚细胞定位预测显示其可能位于质膜上, 多重序列比对及系统进化树结果显示与其他植物GGPPS家族具有较高同源性。实验首次克隆了雷公藤GGPPS 基因,为进一步研究雷公藤甲素等二萜化合物生物合成途径奠定基础。
Abstract:A full-length cDNA of GGPPS gene from Tripterygium wilfordii suspension cells was obtained by use of RACE strategy(GeneBank:KM978333), and then analyzed by bioinformatics approaches. TwGGPPS cDNA has 1857 nucleotides and an open reading frame (ORF) encoding a protein of 514 amino acid residues. The deduced protein has isoelectric point (pI) of 7.85, a calculated molecular weight about 57.13 kD, 5 conserved domains and 2 functional domains. PSORT Prediction showed it was located at plasma membrane. Phylogenetic analysis demonstrated that TwGGPPS1 was similar to GGPPS from other species of plants. For the first time the cloning of geranylgeranyl diphosphate synthase gene from T. wilfordii was reported, it lays the foundation for further research of diterpenoids biosynthetic pathway.
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