津田芜菁BrEGL3 cDNA的克隆及表达分析  

Cloning and Expression Analysis of Enhancer of Glabra3(BrEGL3)in Brassica campestris L. ssp. rapifera‘Tsuda’

作　　者：闫海芳[1] 李玉花[1] 孙佰贺[1] 许道琦[1] 聂唯天[1] 刘振华[1] 闵远琴[1] 

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040

出　　处：《园艺学报》2011年第3期487-494,共8页Acta Horticulturae Sinica

基　　金：国家自然科学基金重点项目(30730078);中央高校基本科研业务费专项(DL09BA09)

摘　　要：EGL3(Enhancer of Glabra3)蛋白是一种bHLH类蛋白,是一类重要的转录调节因子。为阐释BrEGL3在‘津田’芜菁(Brassica campestris L.ssp.rapifera‘Tsuda’)花青素生物合成中的调控作用,根据拟南芥EGL3基因序列信息设计兼并引物,克隆获得了'津田'芜菁EGL3基因的全长cDNA序列,命名为BrEGL3,其GenBank登录号为HM208589。序列分析结果显示,BrEGL3全长1794bp,包含编码597个氨基酸的开放阅读框,分子量约为66.8kD。氨基酸结构分析显示该蛋白含有DNA结合域,该结合属于螺旋-环-螺旋家族。氨基酸对比分析表明,BrEGL3与萝卜EGL3蛋白相似性最高,其次为拟南芥中的EGL3。荧光定量PCR检测BrEGL3在芜菁不同组织中的表达结果表明,该基因在有花青素合成的红色芜菁根皮中表达量最高。Enhancer of Glabra3（EGL3）is a bHLH protein that is an important transcript factor in plant. In order to elucidate the regulation mechanism of EGL3 in‘Tsuda’anthocyanin biosynthesis,cDNA of EGL3 gene was isolated from this plant by the method of RT-PCR using degenerate primers designed according to Arabidopsis EGL3（GenBank accession No. NM202351）. This gene was named BrEGL3 （GenBank accession No. HM208589）. BrEGL3 was 1 794 bp in full length open reading frame（ORF）encoding 597 amino acids with the molecular weight of 66.8 kD. Software prediction results showed that BrEGL3 contained a DNA helix-loop-helix binding domain. Homology analysis showed that the deduced amino acid sequence of BrEGL3 shared the highest similarity with Raphanus sativus EGL3 and the lowest similarity with Lotus angustissimus EGL3. Quantitative-PCR analysis showed that the BrEGL3 was the highest expressed in the in red root epidermis of turnip.

关 键 词：芜菁 EGL3 基因克隆 表达分析 

分 类 号：S631.3[农业科学—蔬菜学]