华东师范大学学报(自然科学版) ›› 2009, Vol. 2009 ›› Issue (2): 96-104.

• 生命科学 • 上一篇    下一篇

一个拟南芥C2H2锌指蛋白的结构域分析

唐洪波，杨扬，孙越，张亮，王洋，李小方，许玲
  

  1. 华东师范大学 生命科学学院,上海200062
  • 收稿日期:2008-09-12 修回日期:2008-12-01 出版日期:2009-03-25 发布日期:2009-03-25
  • 通讯作者: 孙越

Domain function analysis of an Arabidopsis C2H2 zinc finger protein（Chinese）

TANG Hong-bo,YANG Yang,SUN Yue,ZHANG Liang,
WANG Yang,LI Xiao-fang,XU Ling   

  1. School of Life Science, East China Normal University, Shanghai 200062, China
  • Received:2008-09-12 Revised:2008-12-01 Online:2009-03-25 Published:2009-03-25
  • Contact: SUN Yue

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摘要： At3g23140是拟南芥中仅含有一个C2H2锌指结构的转录因子，主要含有N端的C2H2锌指结构和C端的类似EAR两个结构域.将〖WTBX〗〖STBX〗At3g23140〖WTBZ〗〖STB3〗中仅含C2H2锌指结构区域不同长度的两个基因片段〖WTBX〗〖STBX〗A1〖WTBZ〗〖STB3〗(240 bp)，〖WTBX〗〖STBX〗A2〖STB3〗〖WTBZ〗(410 bp)克隆到植物表达载体pMON530 35S启动子的下游，并转化野生型拟南芥.经过筛选得到稳定遗传的T3代纯合转化子.对转基因植株研究表明， 35S::〖WTBX〗〖STBX〗A2〖STB3〗〖WTBZ〗转基因植株的内源乙烯释放量明显低于野生型，这与〖WTBX〗〖STBX〗At3g23140〖STB3〗〖WTBZ〗插入失活突变体〖WTBX〗〖STBX〗cs16966〖STB3〗〖WTBZ〗的表型是一致的，而35S::〖WTBX〗〖STBX〗A1〖WTBZ〗〖STB3〗转基因植株的内源乙烯释放量与野生型没有明显区别.表明A2片段的过量表达产生了显性抑制的作用，这种显性抑制效应很可能是由于 A2蛋白片段与〖WTBX〗〖STBX〗At3g23140〖STB3〗〖WTBZ〗表达产物所调控的核酸序列竞争结合所导致.而A2片段C端所含有的非锌指结构域是At3g23140蛋白与靶基因序列结合所必需的.

关键词: 拟南芥, 锌指蛋白, 转录因子, 竞争抑制, 拟南芥, 锌指蛋白, 转录因子, 竞争抑制

Abstract: At3g23140 is a transcription factor with one single C2H2 zinc finger domain, containing C2H2 zinc finger domain in Nterminal regions and a Cterminal EARmotiflike sequence. Two different fragments of 〖WTBX〗〖STBX〗At3g23140〖STB3〗〖WTBZ〗 that only contain the C2H2type zincfinger domain were inserted downstream of 35S promoter in the plant expression vector pMON530, and introduced into wildtype Landsberg erecta (Ler). Independent homozygous transgenic lines were obtained after selection of T3 progenies. Ethylene detection results showed that endogenous ethylene value of 35S::〖WTBX〗〖STBX〗 A2〖STB3〗〖WTBZ〗 transgenic plants was less than that of the control, which was consistent with the phenotype of〖WTBX〗〖STBX〗 cs16966〖STB3〗〖WTBZ〗, but endogenous ethylene value of transgenic plant 35S::〖WTBX〗〖STBX〗 A1〖STB3〗〖WTBZ〗 has no significant difference with that of the wildtype. It indicated that repression was caused by overexpression of A2, which may due to A2 competing for the DNA sequence regulated by 〖WTBX〗〖STBX〗At3g23140〖STB3〗〖WTBZ〗, and nonC2H2 zinc finger domain of the Cterminal regions of A2 is essential for At3g23140 integrating with targeted DNA.

Key words: zinc finger protein, transcription factor, competition , Arabidopsis thaliana, zinc finger protein, transcription factor, competition

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