摘要
3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)是MVA萜类合成途径的第一个关键酶,在烟草萜类物质的生物合成过程中起到重要的作用.本研究基于转录组数据,从烟草幼叶中克隆出1815 bp、可编码604个氨基酸的HMGR 1基因完整开放阅读框(ORF).经生物信息学分析显示,该基因编码的蛋白序列与番茄相似性最高;该蛋白序列无信号肽,2个区域有跨膜结构域,结构特征为膜外非分泌不稳定疏水蛋白;蛋白质二级结构由α螺旋、延伸链、β转角和不规则卷曲组成,三级结构为同源四聚体.与此同时,运用Real-time PCR对NtHMGR 1基因在烟草不同组织的表达差异性进行检测,结果表明,根<茎<上部叶<中部叶<下部叶.经SA,MEJA,ABA,GA处理后,NtHMGR 1基因的表达峰值出现时间以及在各时间点波动均存在差异性.
3-hydroxy-3-methylglutary Co A reductase(HMGR)is the first key enzyme of the MVA terpenoids synthetic path way,thus it plays an important role in the biosynthesis of Nicotiana tabacum terpenoid.Based on transcriptome data,this experiment cloned the complete open reading frame(ORF)of tobacco HMGR 1 gene from tobacco young leaves by RT-PCR.The gene is 1815 bp in length and encodes 604 amino acids.A comprehensive bioinformatics analysis of the encoded amino acid sequence showed that the protein sequence encoded by this gene is most similar to tomato.The protein sequence does not contain a signal peptide which is a typical unstable hydrophobin and there are two regions that have the transmembrane domain.The secondary structure of its protein sequence is mainly composed ofα-helix and random coil,and the tertiary structure is a homologous tetramer.At the same time,real-time PCR is used to detect the expression of NtHMGR 1 gene in different tissues of tobacco plants.The results showed that the order is root<stem<upper leaf<middle leaf<lower leaf.After treating by SA,MEJA,ABA and GA,there are differences in the peak time of NtHMGR 1 gene expression and the fluctuation trend at each sampling time.
作者
刘晨
王祯
朱先约
杨瑞
张霞
LIU Chen;WANG Zhen;ZHU Xian-yue;YANG Rui;ZHANG Xia(Technology R&D Center,Gansu Tobacco Industry Co.,LTD.,Lanzhou 730050,Gansu,China)
出处
《西北师范大学学报（自然科学版）》
CAS
北大核心
2020年第5期71-77,89,共8页
Journal of Northwest Normal University(Natural Science)
基金
中烟实业科技项目(ZYSY-2019-12)。
