石蒜儿茶酚氧位甲基转移酶基因克隆与原核表达
DOI:
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金(30700057)


Cloning and Prokaryotic Expression of LrCOMT from Lycoris radiata
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    采用同源克隆与RACE相结合的方法,首次从石蒜叶片中克隆到可能与加兰他敏生物合成相关的儿茶酚氧位甲基转移酶基因,命名为LrCOMT。核酸序列分析显示,该cDNA全长1 246 bp,开放阅读框1 101 bp,编码366氨基酸。氨基酸序列分析与比对发现,LrCOMT编码的氨基酸序列具有COMT蛋白的特征区域,且与鸢尾、玉米、烟草等植物COMT的同源性大于60%。将LrCOMT基因连接到原核表达载体pET-29a上,转化大肠杆菌BL21(DE3)的原核表达结果显示,重组蛋白受IPTG诱导表达,分子量大小约为40 kD,与已报道的其他植物COMT大小基本一致。

    Abstract:

    A full-length cDNA of COMT,which is closely related to the biosynthesis of galanthamine in Amaryllidacea plants,was first successfully cloned from Lycoris radiata by the method of homology cloning and RACE,and was named as LrCOMT.The nucleotide analysis showed that the cDNA sequence of LrCOMT gene had 1 246 bp,containing a 1 101 bp open reading frame,which encodes 366 amino acid residues.Amino sequence analysis demonstrated that the deduced amino sequence of LrCOMT has many typical features of COMTs,and shared more than 60% indentity with COMTs from Iris hollandica,Zea mays,Nicotiana tabacum and other plants.Subsequently,the LrCOMT was ligated into pET-29a vector,and transferred into E.coli strain BL21 for heterologous expression.And the result demonstrated that the recombinant protein was induced by IPTG,and its molecular weight is about 40 kD,which is consistent with other COMTs from other plants.

    参考文献
    相似文献
    引证文献
引用本文

梁丽建,江玉梅,夏 冰,等.石蒜儿茶酚氧位甲基转移酶基因克隆与原核表达[J].西北植物学报,2012,32(1):23-28

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2013-01-03
  • 出版日期:

微信公众号二维码

手机版网站二维码