冬凌草HMGS基因的克隆与表达分析  

Molecular Cloning and Expression Analysis of a Gene Encoding 3-Hydroxy-3-Methylglutaryl-CoA Synthase from Isodon rubescen

作　　者：朱畇昊 爼梦航 苏秀红[1,2] 董诚明[1,2] 陈随清[1,2] 

机构地区：[1]河南中医学院药学院,河南郑州450046 [2]呼吸疾病诊疗与新药研发河南省协同创新中心,河南郑州450046

出　　处：《作物杂志》2016年第5期25-30,共6页Crops

基　　金：国家自然科学基金(81173486);河南中医学院创新人才项目(2011XCXRC02);河南省高等学校青年骨干教师计划项目(2011GGJS-089)

摘　　要：羟甲基戊二酰辅酶A合酶(3-Hydroxy-3-methylglutaryl-coenzyme A synthase,HMGS)是甲羟戊酸途径(MVA)中的第一个催化酶。根据冬凌草转录组数据库中HMGS基因序列设计特异引物,采用RT-PCR技术克隆冬凌草IrHMGS基因cDNA全长,并对其序列进行生物信息学分析,通过荧光定量PCR的方法分析其组织表达特性。IrHMGS基因cDNA全长1 382bp,编码460个氨基酸,序列分析结果表明该基因编码的氨基酸序列含有羟甲基戊二酰辅酶A合成酶N末端、C末端等保守结构域。荧光定量PCR表明,IrHMGS在冬凌草组培苗叶和根中的表达量显著高于在组培苗花、茎和愈伤组织中的表达量。本研究为后续深入研究IrHMGS在冬凌草二萜类物质合成途径中的功能奠定了基础。3-hydroxy-3-methylglutaryl coenzyme A synthase （HMGS） catalyzes the first step in the mevalonate biosynthesis pathway in plants. The specific primers were designed according to the transcript sequence of HMGS from the Isodon rubescens transcriptome database. RT-PCR trail was used to clone the full-length cDNA. IrHMGS expression profiles in different tissues were analyzed by quantitative real-time PCR. The full-length cDNA oflrHMGS was found to contain an open reading frame of 1 382bp encoding a polypeptide of 460 amino acid residues. Multiple sequence alignment of IrHGMS with some homologous HMGSs from other plants revealed the IrHMGS had several high conservation domains, such as Hydroxymethylglutaryl-coenzyme A synthase N-terminal, C-terminal domain. Expression profile analysis revealed that the IrHGMS expression level in leaves and roots was higher than that in flowers, stems and calluses of I. rubescens. The characterization and expression provides useful information for further studying this gene and its function in the diterpene biosynthetic pathway in I. rubescens.

关 键 词：冬凌草 HMGS基因 生物信息学分析 荧光定量PCR 

分 类 号：S567.239[农业科学—中草药栽培]