石斛兰dfr基因的克隆、序列分析及原核表达  

Cloning,Sequencing and Prokaryotic Expression of dfr from Dendrobium

作　　者：潘丽晶[1] 张妙彬[1] 范干群[1] 陈伟庭[1] 曹友培[1] 

机构地区：[1]珠海市农业科学研究中心,广东珠海519070

出　　处：《园艺学报》2010年第1期129-134,共6页Acta Horticulturae Sinica

基　　金：广东省科技计划项目(2006B20130004)

摘　　要：二氢黄酮醇4-还原酶(DFR)在不同花色形成中起着关键作用。利用RT-PCR方法从石斛兰中克隆出dfr基因,并命名为Dendfr。该序列全长1164bp,编码352个氨基酸。氨基酸序列分析发现,DenDFR与3种兰科植物的DFR氨基酸序列同源性达81%～86%,与其他非兰科植物的DFR氨基酸同源性在56%～72%之间。在DenDFRN-末端存在一个保守的NADP结合区,序列中存在26个氨基酸组成的底物特异结合区,其中134位氨基酸处存在特异的天冬酰胺N位点。将Dendfr的编码区克隆到pQE30载体中,在大肠杆菌中高效表达了该蛋白,为进一步研究石斛兰花色形成机制及花色改良基因工程打下基础。Dihydroflavonol 4-Reductase（DFR）plays a key role in the formation of different floral color.By using RT-PCR method,a dfr gene was cloned from Dendrobium and designated as Dendfr.The Dendfr gene was 1 164 bp in length and encoded 352 amino acid residues.Amino acid sequence analysis revealed that Den DFR sequence share homologies of 81%-86% with three species of orchid,and share homologies of 56%-72% with the other species.Den DFR protein contains a reserved NADP domain at the N-terminal and a specific 26 amino acid region of substrate specificity with an asparagine（position 134）.The encoding region of Dendfr was constructed into pQE30 vector and effectively expressed the protein in E.coli.These will be helpful for studying molecular mechanism of floral color formation and genetic improvement of the Dendrobium.

关 键 词：石斛兰 花色 dfr基因 克隆 序列分析 原核表达 

分 类 号：S682.31[农业科学—观赏园艺] Q78[农业科学—园艺学]