天然橡胶合成关键酶基因Hmg1启动子克隆与序列分析  

Cloning, Sequence Analysis and Expression Vector Construction of 5' Regulation Region for Key Enzyme Gene of Natural Rubber Biosynthesis from Hevea brasiliensis

作　　者：张福城[1] 邓治[1] 张云霞[1] 陈守才[1] 

机构地区：[1]中国热带农业科学院橡胶栽培研究所农业部热带作物栽培生理学重点开放实验室,海南儋州571737

出　　处：《热带作物学报》2006年第3期40-45,共6页Chinese Journal of Tropical Crops

基　　金：973重大基础研究前期研究专项(2004CCA00400)资助

摘　　要：在巴西橡胶树天然橡胶的生物合成途径中,3-羟基-3-甲基戊二酸单酰辅酶A还原酶(3-Hydroxy-3-MethylglutarylCoenzymeAReductase,HMGR)是关键酶之一,为了利用其基因的启动子和瞬时表达系统来筛选促进橡胶树生物合成的调节剂,利用GenomicWalking方法从高产树热研7-33-97的叶片基因组DNA中克隆了Hmg15′端调控序列。序列分析结果表明,巴西橡胶树Hmg1启动子转录起始位点的保守序列CTCATCA,与其他植物基因的非常一致;在5′端调控序列中发现几个典型的真核生物顺式调控元件TATA-motif、CAAT-motif、CGTCA-motif、ERE等。用克隆得到的Hmg15′端启动子序列分别替换了pCAMBIA1302中的CaMV35S启动子,构建了序列缺失pCAMBIA1302表达载体pCAMBIA1302H900、pCAMBIA1302H500和pCAMBIA1302H300。The 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase was one of the key enzymes in the rubber biosynthesis pathway. For screening the regulator of promoting rubber biosynthesis by using the Hmg1 Promoter and instantaneous expression system, the 5＇regulation region for Hmg1 was obtained using Genomic Walking from the leaf genome DNA of high production Hevea brasiliensis 7-33-97. Sequence analysis demonstrated that the conservative sequence CTCATCA at the transcription start site of Hmg1 gene was quite similar to that of other plant genes. Meanwhile, several typical eukaryotic cis-regulatory elements （TATA-motif, CAAT-motif, CGTCA- motif, ERE and so on） were found in the 5＇ regulation region. CaMV35S promoter of pCAMBIA1302 was replaced with the three cloned fragments, and the deletion expression and pCAMBIA1302H300 were constructed. vectors of pCAMBIA1302H900 pCAMBIA1302H500

关 键 词：巴西橡胶树 Hmg1启动子 克隆与分析 表达载体构建 

分 类 号：S794.1[农业科学—林木遗传育种]