| 酿酒酵母钙调蛋白依赖性激酶-2的过量表达及其在氧化胁迫应答中的作用初探 |
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| 引用本文： | 薛薇,孙毅,钟彦,朱德燕,刘科. 酿酒酵母钙调蛋白依赖性激酶-2的过量表达及其在氧化胁迫应答中的作用初探[J]. 四川大学学报(自然科学版), 2009, 46(5): 1553-1557. DOI: 103969/j.issn.0490-6756.2009.05.64 |
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| 作者姓名： | 薛薇 孙毅 钟彦 朱德燕 刘科 |
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| 作者单位： | 四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都,610064;四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都,610064;四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都,610064;四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都,610064;四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都,610064 |
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| 摘 要： | 利用PCR方法扩增钙调蛋白依赖性激酶（CaM-dependent kinase ,cmk）-2基因的蛋白质编码序列,经限制性内切酶Kpn I和BamH I酶切后连接入空载体pYES2/NTA构建真核表达载体,构建的pYES2/NTA-cmk2转化酿酒酵母菌株BY4741进行真核表达.经诱导和收集细胞后,对酵母细胞用玻璃珠震荡破壁,收集上清液进行镍离子亲和层析,并纯化Cmk2,纯化的蛋白通过Western blot检测.并且初步研究了Cmk2在酵母细胞氧化胁迫应答中的作用.
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| 关 键 词： | 钙调蛋白依赖性激酶 酿酒酵母 过量表达 氧化胁迫应答 |
| 收稿时间： | 2008-04-18 |
| 修稿时间： | 2008-05-28 |
Overexpression of CaM-dependent kinase-2 (Cmk2) and initiate study on its role in oxidative stress response in Saccharomyces cerevisiae |
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| XUE Wei,SUN Yi,ZHONG Yan,ZHU De Yan and LIU Ke. Overexpression of CaM-dependent kinase-2 (Cmk2) and initiate study on its role in oxidative stress response in Saccharomyces cerevisiae[J]. Journal of Sichuan University (Natural Science Edition), 2009, 46(5): 1553-1557. DOI: 103969/j.issn.0490-6756.2009.05.64 |
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| Authors: | XUE Wei SUN Yi ZHONG Yan ZHU De Yan LIU Ke |
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| Affiliation: | Key Laboratory of Bio-resource and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University;Key Laboratory of Bio-resource and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University;Key Laboratory of Bio-resource and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University;Key Laboratory of Bio-resource and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University;Key Laboratory of Bio-resource and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University |
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| Abstract: | The ORF of CaM-dependent kinase-2 was amplified by PCR, and cloned into expression vector of pYES2/NTA to construct recombinant expression plasmid pYES2/NTA-cmk2. The recombinant plasmid was transformed into Saccharomyces cerevisiae strain BY4741 and overexpressed. The His6 tagged fusion protein was purified by Ni2+ affinity chromatography and further analyzed by SDS-PAGE and Western blot. Role of Cmk2 in oxidative stress response was studied initiatively. |
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| Keywords: | CaM-dependent kinase-2 Saccharomyces cerevisiae overexpression oxidative stress response |
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