中药材2017，Vol.40Issue(8)：1793-1797,5.DOI:10.13863/j.issn1001-4454.2017.08.010
艾叶1-羟基-2-甲基-2-(E)-丁烯基-4-焦磷酸还原酶基因的克隆及蛋白的结构性质分析
Cloning and Characterization of 1-Hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate Reductase (ArHDR) Gene in Artemisia argyi
摘要
Abstract
Objective:To clone the open reading frame (ORF)of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (ArHDR) from Artemisia argyi,and to analyze the characteristics and structure of the coding protein.Methods:The target gene fragments were amplified by reverse transcriptase PCR method and subcloned to pMD19-T vectors,and then the recombinant plasmids were transformed into Escherichia coli DH5α cells.The positive clones were confirmed by bacteria PCR and sequencing.The coding protein ArHDR was analyzed by systematic bioinformatics softwares.Results:The full-length ORF of the ArHDR,which contained 1 365 bp encoding ArHDR protein with 454 amino acids,was acquired.Bioinformatics analysis showed that ArHDR was a stable protein and it has the closest relationship with HDR from Artemisia annua.ArHDR showed cloverleaf structure consisting of three lobes,each containing alternating α/β structure.Three conserved cysteines located in the center of the three dimensional structure,which formed the catalytic region for anchoring iron-sulfur.Conclusion:The ArHDR is successfully cloned in vitro,and the property and structure of ArHDR are studied.The results provide theoretical basis for promoting the synthesis of terpenoids from Artemisia argyi.关键词
艾叶/ArHDR基因/基因克隆/生物信息学分析Key words
Artemisiae argyi Lévl.et Vant./ArHDR gene/Gene clone/Bioinformatics analysis分类
医药卫生引用本文复制引用
刘苗苗,吴家文,吴生兵,杨青山,周美启..艾叶1-羟基-2-甲基-2-(E)-丁烯基-4-焦磷酸还原酶基因的克隆及蛋白的结构性质分析[J].中药材,2017,40(8):1793-1797,5.基金项目
安徽高校科研创新平台团队建设项目(2015TD033) （2015TD033）
安徽省自然科学基金(1608085MH177) （1608085MH177）
安徽省高校优秀中青年骨干人才国内外访学研修重点项目(gxfxZD2016115) （gxfxZD2016115）
安徽中医药大学人才引进科研项目(2015RC002) （2015RC002）
安徽省留学人员科技活动启动项目(2015lx024) （2015lx024）
