食品科学 ›› 2010, Vol. 31 ›› Issue (13): 219-223.doi: 10.7506/spkx1002-6630-201013050

• 生物工程 • 上一篇    下一篇

苦荞和甜荞二氢黄酮醇4-还原酶基因(Dfr)的克隆及序列分析

祝 婷,李成磊,吴 琦﹡,蒙 华,陈 惠,邵继荣   

  1. 四川农业大学生命科学与理学院
  • 收稿日期:2009-12-30 修回日期:2010-04-12 出版日期:2010-07-01 发布日期:2010-12-29
  • 通讯作者: 吴琦 E-mail:wuqiwq@yahoo.cn
  • 基金资助:

    四川省科技厅科技攻关项目(04NG001-015;2006Z08-012)

Cloning and Sequence Analysis of Dihydroflavonol 4-Reductase Genes from Tartary Buckwheat and Common Buckwheat

ZHU Ting,LI Cheng-lei,WU Qi*,MENG Hua,CHEN Hui,SHAO Ji-rong   

  1. College of Life Science, Sichuan Agricultural University, Ya’an 625014, China
  • Received:2009-12-30 Revised:2010-04-12 Online:2010-07-01 Published:2010-12-29
  • Contact: WU Qi*, E-mail:wuqiwq@yahoo.cn

摘要:

采用同源基因克隆的方法,以苦荞和甜荞叶片为材料,提取总RNA,经RT-PCR 扩增获得其二氢黄酮醇4- 还原酶基因(dfr)cDNA 序列,并通过T 载体克隆后测序。序列分析表明,获得了苦荞和甜荞dfr 的全长cDNA编码序列,其1026bp 的全长开放阅读框(ORF)均编码341 个氨基酸,并具典型的dfr 结构特征和功能模块。同源性分析显示,苦荞和甜荞与其他植物的dfr 基因核苷酸相似性为71%~98%;根据氨基酸序列构建系统进化树表明,二者与豆科、桑科和蔷薇科聚为一类。

关键词: 苦荞, 甜荞, 二氢黄酮醇4 - 还原酶基因, 克隆, 序列分析

Abstract:

The cDNA sequence of dihydroflavonol 4-reductase (DFR) gene was amplified from total RNA from the leaves of tartary buckwheat (Fagopyrum tataricum) and common buckwheat (Fagopyrum esculentum) by RT-PCR using homology-based cloning strategy. The amplified fragments were then cloned into T-vector. Sequence analysis indicated that both cDNAs had open reading frames of 1026 bp in full length, which encoded a polypeptide composed of 341 amino acids with typical structure characteristics and functional module of DFR enzyme, respectively. The nucleotide similarity of dfr gene among Fagopyrum tataricum, Fagopyrum esculentum and other plants ranged from 71% to 98%. In addition, the phylogenetic analysis based on amino acid sequence enconded by dfr gene indicated that both buckwheat varieties, Leguminosae, Moraceae and Rosaceae were classified into one class.

Key words: Fagopyrum tataricum Gaertn, Fagopyrum esculentum Moench, dihydroflavonol 4-reductase (dfr) gene, cloning, sequence analysis

中图分类号: