三色堇4,5-多巴雌二醇双加氧酶基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of 4,5-DOPA Dioxygenase Extradiol Gene in Viola wittrockiana

作　　者：李霆格 杨文汉[1] 曾媛[1] 李婧[1] 赵安瑾 王健[1] Li Tingge Yang Wenhan Zeng Yuan Li Jing Zhao Anjin Wang Jian(Key Laboratory of Protection and Development Utilization of Tropical Crop Germplasm Resources, College of Horticulture ＆ Landscape Architec-ture, Hainan University, Haikou, 57022)

机构地区：[1]海南大学园艺园林学院,热带作物种质资源保护与开发利用教育部重点实验室,海口570228

出　　处：《分子植物育种》2017年第2期433-440,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31060265;31260488);海南大学中西部计划学科建设项目(ZXBJH-XK008)共同资助

摘　　要：本实验以三色堇(Viola wittrockiana)的花瓣为研究材料,提取其总RNA,通过RACE(rapid-amplification of cDNA ends)技术扩增4,5-多巴雌二醇双加氧酶(4,5-DOPA dioxygenase extradiol)基因的cDNA全长,并对其进行生物信息学分析。本实验成功克隆VwDODA的cDNA全长1 055 bp,其中3'端非编码区157 bp,包含29 bp Poly(A),5'端非编码区103 bp,其开放阅读框为795 bp,编码263个氨基酸。通过NCBI的BLAST比对表明VwDODA编码的氨基酸序列与其他植物相关基因的相似性达到80.29%,与大豆(Glycine max)亲缘关系最近。通过生物信息学分析软件对VwDODA的蛋白质结构与功能进行预测,结果表明Vw D-%ODA是一种亲水稳定蛋白,不存在跨膜区域和信号肽,分子式为C1337H2026N356O384S7,等电点为6.70,分子量为29 455.37。本研究为将来三色堇中甜菜色素与花色素的互斥现象研究打下了基础。In this research, the full-length of cDNA of 4,5-DOPA dioxygenase extradiol of Viola wittrockiana was cloned by the method of RACE using the petals as the materials, and the bioinformatics analysis of this gene was conducted. The length of cDNA of the cloned%VwDODA was 1 055 bp, with 157 bp length of 3＇ untranslated region containing 29 bp Poly（A）, and 103 bp length of 5＇ untranslated region. The open read frame was 795 bp, encoding263 amino acids. The Blast revealed that the amino acids sequence encoded by VwDODA had 80.29% consistency compared to the same genes of other plants, and the genetic relationship was close to Glycine max. The structure prediction of VwDODA found that it was a kind of hydrophilic and stable protein with no transmembrane region or signal peptide, of which the molecular formula was C1337H2026N356O384S7, and the isoelectric point was 6.70 and the molecular weight was 29 455.37. This study had laid a solid foundation that could reveal the mutually exclusive nature of the betalain and anthocyanin in pansy or even other plants.

关 键 词：三色堇(Viola wittrockiana) 4 5-多巴雌二醇双加氧酶(DODA) cDNA末端快速克隆(RACE) 生物信息学 

分 类 号：S682.19[农业科学—观赏园艺] Q943.2[农业科学—园艺学]