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| 普通小麦中双脱氢抗坏血酸还原酶(TaDHAR)基因的克隆与生化特性分析 | |
| 引用本文： | 余春梅,杨艳萍,刘鑫燕,周蓉,华梁,魏贺,丁胜杰,王道文.普通小麦中双脱氢抗坏血酸还原酶(TaDHAR)基因的克隆与生化特性分析[J].生物工程学报,2009,25(10):1483-1489. |
| 作者姓名： | 余春梅 杨艳萍 刘鑫燕 周蓉 华梁 魏贺 丁胜杰 王道文 |
| 作者单位： | 1. 南通大学生命科学学院,南通,226007 2. 中国科学院遗传与发育生物学研究所,北京,100101;中国科学院研究生院,北京,100049 3. 中国科学院遗传与发育生物学研究所,北京,100101 |
| 基金项目： | 国家自然科学基金项目(No.30771306);;南通大学博士科研基金项目(No.09B04)资助~~ |
| 摘 要： | 利用同源克隆技术从六倍体普通小麦中获得了两个不同的双脱氢抗坏血酸还原酶(TaDHAR)基因的cDNA克隆。器官表达模式分析表明,这两个TaDHAR基因(暂时命名为TaDHAR1和TaDHAR2)在小麦根、茎、叶、幼穗以及开花后10d、20d和30d的种子中均有表达,为组成型表达基因。原生质体表达实验表明,两个基因的产物均可能定位在细胞质中。在细菌中表达并提纯了两个基因的重组蛋白。体外生化测定表明两个重组蛋白均具有将双脱氢抗坏血酸还原成抗坏血酸的能力,其最适pH为7.5,在37oC时的活性比25oC高,但25oC条件下pH6.0和7.0时,两个DHAR蛋白的活性显著不同。本研究的结果为进一步揭示TaDHAR基因在小麦抗坏血酸代谢中的生理作用奠定了基础。 |
| 关 键 词： | 普通小麦 双脱氢抗坏血酸还原酶 抗坏血酸 双脱氢抗坏血酸 表达模式 酶活 |
| 收稿时间： | 2009/5/13 0:00:00 |
Molecular and biochemical analysis of two genes encoding dehydroascorbate reductase in common wheat |
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| Chunmei Yu,Yanping Yang,Xinyan Liu,Rong Zhou,Liang Hu,He Wei,Shengjie Ding and Daowen Wang.Molecular and biochemical analysis of two genes encoding dehydroascorbate reductase in common wheat[J].Chinese Journal of Biotechnology,2009,25(10):1483-1489. | |
| Authors: | Chunmei Yu Yanping Yang Xinyan Liu Rong Zhou Liang Hu He Wei Shengjie Ding and Daowen Wang |
| Institution: | School of Life Sciences, Nantong University, Nantong 226007, China;Institute of Genetics and Developmental Biology, Chinese Academy of Science, Beijing 100101, China; Graduate University of Chinese Academy of Sciences, Beijing 100049, China;School of Life Sciences, Nantong University, Nantong 226007, China;School of Life Sciences, Nantong University, Nantong 226007, China;School of Life Sciences, Nantong University, Nantong 226007, China;School of Life Sciences, Nantong University, Nantong 226007, China;School of Life Sciences, Nantong University, Nantong 226007, China;Institute of Genetics and Developmental Biology, Chinese Academy of Science, Beijing 100101, China |
| Abstract: | Dehydroascorbate reductase (DHAR) plays an important role in the recycling of ascorbic acid. In this work, we isolated the full length cDNA clones of two different DHAR genes (tentatively named as TaDHAR1 and TaDHAR2, respectively) from common wheat. Semi-quantitative PCR experiments showed that TaDHAR1 and TaDHAR2 were transcribed in many vegetative and reproductive organs examined in this work. Transient expression analysis using wheat protoplasts indicated that the protein products of TaDHAR1 and TaDHAR2... |
| Keywords: | common wheat dehydroascorbate reductase ascorbic acid dehydroascorbate expression pattern enzyme activity |
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