天然产物研究与开发 ›› 2016, Vol. 28 ›› Issue (10): 1505-1509.doi: 10.16333/j.1001-6880.2016.10.001

• 研究论文 •    下一篇

川芎咖啡酸-O-甲基转移酶基因的克隆及序列分析

朱建全,向缅,李洋洋,俞继华,张赶,王万军,廖海*,周嘉裕*   

  1. 西南交通大学生命科学与工程学院,成都 610031
  • 出版日期:2016-10-31 发布日期:2017-03-30

Molecular Cloning and Sequence Analysis of Caffeic Acid -O-Methyltransferase in Ligusticum chuanxiong Hort

ZHU Jian-quan,XIANG Mian,LI Yang-yang,YU Ji-hua,ZHANG Gan,WANG Wan-jun,LIAO Hai*,ZHOU Jia-yu*   

  1. School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031,China
  • Online:2016-10-31 Published:2017-03-30

摘要: 克隆川芎(Ligusticum chuanxiong Hort)咖啡酸-O-甲基转移酶(COMT)基因并做序列分析。采用RT-PCR方法,以新鲜的川芎嫩叶cDNA为模板,克隆出川芎COMT基因的cDNA序列。此基因全长1328 bp,编码360个氨基酸。将得到的序列提交GenBank,序列号为KU942388。此川芎COMT具有植物-O-甲基转移酶家族特征性的功能结构域,与中粒咖啡、牵牛花COMT的氨基酸序列同源性分别为70%、69%,推测其能够催化咖啡酸的氧甲基化反应。该川芎COMT基因的成功克隆为下一步活性验证及利用生物技术来生产阿魏酸奠定坚实基础。

关键词: 川芎, 咖啡酸-O-甲基转移酶, 克隆, 序列分析

Abstract: In order to clone and sequence the cDNA encoding caffeic acid-O-methyltransferase (COMT) from Ligusticum chuanxiong Hort,the cDNA,encoding COMT,was amplified by RT-PCR with cDNA library of rhizome as the template.The full-length cDNA of COMT was 1328 bp with an open reading frame encoding 360 amino acids of protein.The accession number of COMT from L.chuanxiong in GenBank was KU942388.The COMT from L.chuanxiong has 70% and 69% nucleotide sequence homology to the sequence of COMT from Coffea canephora and Ipomoea nil, respectively.Comparison of the sequence L.chuanxiong with COMT from C.canephora and I.nil showed that COMT from L.chuanxiong possessed the same functional regions involved with COMT,suggesting L.chuanxiong COMT can participate in oxygen methylation of caffeic acid.This work underlaid the first step for using biotechnology to produce ferulic acid.

Key words: Ligusticum chuanxiong Hort, caffeic acid-O-methyltransferase, gene cloning, sequence analysis

中图分类号: 

Q785