中国中药杂志

2015, v.40(06) 1066-1070

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雷公藤牻牛儿基牻牛儿基焦磷酸合酶基因全长cDNA的获得及生物信息学分析
Cloning and bioinformatics analysis of geranylgeranyl diphosphate synthase gene of Tripterygium wilfordii

张萌;苏平;刘雨佳;童宇茹;赵瑜君;高伟;王秀娟;黄璐琦;
ZHANG Meng;SU Ping;LIU Yu-jia;TONG Yu-ru;ZHAO Yu-jun;GAO Wei;WANG Xiu-juan;HUANG Lu-qi;School of Traditional Chinese Medicine,Capital Medical University;National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;

摘要(Abstract):

该研究利用c DNA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c DNA(Gene Bank: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Tw GGPPS c DNA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 k Da,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。
A full-length c DNA of GGPPS gene from Tripterygium wilfordii suspension cells was obtained by use of RACE strategy( Gene Bank: KM978333),and then analyzed by bioinformatics approaches. Tw GGPPS c DNA has 1857 nucleotides and an open reading frame( ORF) encoding a protein of 514 amino acid residues. The deduced protein has isoelectric point( p I) of 7. 85,a calculated molecular weight about 57. 13 k D,5 conserved domains and 2 functional domains. PSORT Prediction showed it was located at plasma membrane. Phylogenetic analysis demonstrated that Tw GGPPS1 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.

关键词(KeyWords): 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
Tripterygium wilfordii;GGPPS;clone;bioinformatics analysis

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基金项目(Foundation): 国家自然科学基金项目(81422053,81325023);; 北京市属高等学校高层次人才引进与培养计划项目(CIT&TCD201304174);; 全国优秀博士学位论文作者专项资金项目(201188)

作者(Author): 张萌;苏平;刘雨佳;童宇茹;赵瑜君;高伟;王秀娟;黄璐琦;
ZHANG Meng;SU Ping;LIU Yu-jia;TONG Yu-ru;ZHAO Yu-jun;GAO Wei;WANG Xiu-juan;HUANG Lu-qi;School of Traditional Chinese Medicine,Capital Medical University;National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;

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