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中国科学院大学学报 ›› 2013, Vol. 30 ›› Issue (3): 329-333.DOI: 10.7523/j.issn.1002-1175.2013.03.008

• 化学与生命科学 • 上一篇    下一篇

顺乌头酸酶AtACO3参与调控Zn稳态的研究

李燕, 王建武, 谭金娟, 冯珊珊, 柴团耀   

  1. 中国科学院研究生院生命科学学院, 北京 100049
  • 收稿日期:2012-03-16 修回日期:2012-04-28 发布日期:2013-05-15
  • 通讯作者: 柴团耀, tychai@ucas.ac.cn
  • 基金资助:

    中国科学院知识创新工程重要方向项目——生命科学领域十二五基础前沿专项(KZCX2-EW-J-29)资助

Involvement of AtACO3 in Zn homeostasis

LI Yan, WANG Jian-Wu, TAN Jin-Juan, FENG Shan-Shan, CHAI Tuan-Yao   

  1. College of Life Science, Graduate University, Chinese Academy of Sciences, Beijing 100049, China
  • Received:2012-03-16 Revised:2012-04-28 Published:2013-05-15

摘要:

从拟南芥中克隆得到AtACO3基因的全长cDNA序列. 半定量PCR实验结果表明,200 μmol/L CuCl2处理能明显抑制该基因的表达,而200 μmol/L ZnCl2处理1 h则可显著促进其表达. 为验证AtACO3蛋白N端的功能,构建AtACO3 5'端1632 bp基因片段的原核表达载体pET30a-AtACO3-N,并转化大肠杆菌. SDS-聚丙烯酰胺凝胶电泳检测结果显示,该基因片段可以在大肠杆菌中稳定表达. 进一步的抗性实验表明,异源表达基因AtACO3的N端序列能提高大肠杆菌的Zn2+耐受性.

关键词: Zn, 拟南芥, 顺乌头酸酶

Abstract:

A full-length cDNA, designated as AtACO3, was isolated from Arabidopsis thaliana. Semi-quantitative RT-PCR revealed that AtACO3 was significantly activated under treatment with 200 μmol/L ZnCl2 for 1 hour while inhibited under treatment with 200 μmol/L CuCl2. To investigate the function of the N-terminus of AtACO3, the prokaryotic expression vector pET30a with a fragment of AtACO3 including 1632 bp from the initiation codon was constructed and successfully transformed E. coli. SDS-PAGE indicated that the recombinant N-terminus of AtACO3 was soluble and stable when expressed in E. coli. Additionally, the tolerance of E. coli against Zn2+ was found to be enhanced by expressing the N-terminus fragment of AtACO3.

Key words: Zn, Arabidopsis thaliana, aconitase

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