基于转录组信息的艾纳香牻牛儿基焦磷酸合成酶基因(BbGPPS)的克隆及序列分析  

Cloning and Analysis of Geranyl Pyrophosphate Synthase(GPPS) Sequence of Blumea balsamifera L.DC on Transcriptome Information

作　　者：官玲亮[1] 夏奇峰[1,2] 石小兵[2] 蓝惠萍 陈振夏[1] 赵致[2] 庞玉新[1] 

机构地区：[1]中国热带农业科学院热带作物品种资源研究所,农业部华南作物基因资源与种质创制重点开放实验室,海南省艾纳香工程技术研究中心,海南儋州571737 [2]贵州大学生命科学学院,贵州省药用植物繁育与种植重点实验室,贵州贵阳550025

出　　处：《热带作物学报》2016年第5期901-909,共9页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金(No.81202910);中央级公益性科研院所基本科研业务费专项资金(No.1630032014015);海南省中药现代化专项(No.ZY201410)

摘　　要：采用RT-PCR和RACE(Rapid amplification of cDNA ends)技术,从艾纳香(Blumea balsamifera L.DC)的叶片中克隆到单萜化合物合成的关键酶牻牛儿基焦磷酸合成酶(BbGPPS)基因。研究结果显示,BbGPPS基因的cDNA全长1 692 bp,包含开放阅读框(ORF)1 083 bp,编码361个氨基酸;亚细胞结构定位于叶绿体,既非膜蛋白也非分泌性蛋白。疏水性分析显示,BbGPPS是亲水性蛋白。同源性比对结果显示,BbGPPS蛋白与其他植物中GPPS蛋白具有高度的相似性,且具有异戊烯基结构域。系统发育分析表明,所有序列被聚为5大类,BbGPPS与其他菊科植物聚类一类,与万寿菊(Tagetes erecta)TeGPPS亲缘关系最近,其次是甜菊(Stevia rebaudiana)SrGPPS。A geranyl diphosphate synthase gene, designated BbGPPS, was cloned from Blumea balsamifera L. DC using reverse transcription polymerase chain reaction approach （RT-PCR） and rapid amplification of eDNA ends （RACE） methods. The results showed that The BbGPPS cDNA had a full length of 1 692 bp, and contained an open reading frame predicting a polypeptide of 361 amino acids. Hydropathy and subcellular localization prediction showed that the BbGPPS belonged to hydrophilic protein and located in Chloroplasts. It was neither a membrane protein nor a secretory protein. BbGPPS protein showed a high similarity with other plant GPPS genes, containing isopentenyl domain. Phylogenetic analysis indicated that the amino acid sequence was divided into five categories and BbGPPS grouped with other composite plant GPPS, such as Tagetes erecta TeGPPS and Stevia rebaudiana SrGPPS.

关 键 词：艾纳香 牻牛儿基焦磷酸合成酶 氨基酸序列 系统发育分析 

分 类 号：S567[农业科学—中草药栽培] Q78[农业科学—作物学]