丹参2-酮戊二酸依赖性双加氧酶基因克隆及表达分析  

Cloning and Expression Analysis of 2-oxoglutarate-dependent Dioxygenase from Salvia miltiorrhiza Bunge

作　　者：刘文超[1] 王东浩[1] 王喆之[1] 李翠芹[1] 

机构地区：[1]教育部药用资源与天然药物化学重点实验室,西北濒危药材资源开发国家工程实验室,陕西师范大学生命科学学院,西安710062

出　　处：《西北植物学报》2012年第7期1289-1294,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：陕西师范大学中央高校基本科研业务费重点项目(GK200901014)

摘　　要：以商洛紫花丹参为材料,对其转录组序列SRA020132进行Blast分析,采用PCR技术克隆得到丹参2-酮戊二酸依赖性双加氧酶基因Sm2-ODD1,GenBank登录号为JN935923。Sm2-ODD1基因全长1 365bp,包含3个外显子和2个内含子;cDNA全长1 189bp,读码框951bp,编码316个氨基酸残基;预测的编码蛋白具有2-酮戊二酸依赖性双加氧酶中结合2-酮戊二酸和亚铁离子的"H-T-D"、"H-X"和"R-Y-S"保守基序以及"果冻状"空间结构。表达分析显示,Sm2-ODD1在丹参各个器官都表达,但表达水平具有组织特异性,在根中表达量最高,在叶中表达量最低;该基因表达明显受到MeJA、GA3和ABA的诱导,可能参与了丹参萜类代谢下游途径。By analyzing Transcriptome（SRA020132） sequences of Salvia miltiorrhiza from Shangluo and using the techniques of PCR,a new 2-oxoglutarate-dependent dioxygenase gene was cloned from Salvia miltiorrhiza for the first time and named as Sm2-ODD1（GenBank accession number JN935923）.Sm2-ODD1 DNA consisted of 1 365 bp including 3 exons and 2 introns.The full-length cDNA of Sm2-ODD1 was 1 189 bp,containing a single 951 bp opening reading frame and encoding a 316 amino-acid residues.Bioinformatics analysis showed that Sm2-ODD1 contained a 2-oxoglutarate-dependent dioxygenase superfamily containing ＂distorted jelly roll＂ domain and ＂H-T-D＂,＂H-X＂ and ＂R-Y-S＂ motifs to bind 2-oxoglutarate and Fe2＋.Quantitative RT-PCR analysis revealed that the gene expressed in different organs.The expression in roots was observably higher than in flowers,stems and leaves.The gene could be induced by methyl jasmonate,GA3 and ABA.The results indicated that the Sm2-ODD1 might be involved in the biosynthesis of terpene in Salvia miltiorrhiza.

关 键 词：丹参 2-酮戊二酸依赖性双加氧酶 基因克隆 表达特性分析 实时荧光定量PCR 

分 类 号：Q789[生物学—分子生物学]