| 顺乌头酸酶AtACO3参与调控Zn稳态的研究 |
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| 作者姓名： | 李燕 王建武 谭金娟 冯珊珊 柴团耀 |
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| 作者单位： | 中国科学院研究生院生命科学学院, 北京 100049 |
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| 基金项目： | 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 |
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| 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.
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| 关 键 词： | Zn 拟南芥 顺乌头酸酶 |
| 收稿时间： | 2012-03-16 |
| 修稿时间： | 2012-04-28 |
Involvement of AtACO3 in Zn homeostasis |
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| Authors: | LI Yan WANG Jian-Wu TAN Jin-Juan FENG Shan-Shan CHAI Tuan-Yao |
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| Institution: | College of Life Science, Graduate University, Chinese Academy of Sciences, Beijing 100049, China |
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| 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. |
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| Keywords: | Zn Arabidopsis thaliana" target="_blank">Arabidopsis thaliana')">Arabidopsis thaliana aconitase |
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