花生AhDGAT1 基因启动子的克隆与功能验证

摘要

二酰基甘油酰基转移酶(DGAT)是三酰基甘油(TAG)生物合成过程的限速酶.在本研究中,通过染色体步移法从栽培花生品种鲁花14基因组中克隆了AhDGAT1的启动子(pAhDGAT1)序列,并通过生物信息学分析发现该启动子含有多个 TATA-box、CAAT-box、光调控元件、胁迫防御相关元件和激素响应元件.利用农杆菌介导法将pAhDGAT1∶GUS植物表达载体转化烟草叶片.通过GUS染色发现在转基因烟草营养器官和生殖器官中均检测到GUS表达,在花丝和花柱中表达较低,而在柱头、花药和种子中表达较高,表明pAhDGAT1表达没有组织特异性,具有组成型启动子的特征.%Diacylglycerol acyltransferase (DGAT) is a rate-limiting enzyme in the biosynthesis pathway of triacylglycerol (TAG).In this study, with peanut cultivar Luhua 14 as material, we cloned the promoter sequence of AhDGAT1 gene (pAhDGAT1) by genome walking method.By bioinformatics analysis, we found that the pAhDGAT1 had many TATA-box, CAAT-box, light regulation, stress and defense response and hormone response elements.The constructed pAhDGAT1∶GUS plant expression vector was transformed into tobacco leaves by Agrobacterium tumefaciens-mediated method.By GUS staining,we found the GUS enzyme activity were detected in all organs of the positive transgenic tobacco lines, while in stigma, anther and young seed, the expression levels were higher than those in filament and style.It indicated that pAhDGAT1 was not tissue-specific promoter and had constitutive promoter feature.