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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 被引量：1

Overexpression of CaM-dependent kinase-2(CMK2)and initiate study on its role in oxidative stress response in Saccharomyces cerevisiae
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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 2)基因的蛋白质编码序列,经限制性内切酶KpnI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,纯化的蛋白通过免疫杂交检测.并且初步研究了CMK2在酵母细胞氧化胁迫应答中的作用. 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 Ni^2＋ affinity chromatography and further analyzed by SDS-PAGE and immune blot. Role of CMK2 in oxidative stress response was studied initiatively.
作者 薛薇 孙毅 钟彦 朱德燕 刘科
机构地区 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
出处 《四川大学学报（自然科学版）》 CAS CSCD 北大核心 2009年第5期1553-1557,共5页 Journal of Sichuan University(Natural Science Edition)
基金 国家自然科学基金(30671181)
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 CaM-dependent kinase-2, Saccharomyces cerevisiae, overexpression, oxidative stress response
分类号 Q81 [生物学—生物工程]
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参考文献10

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共引文献1

同被引文献23

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四川大学学报（自然科学版）
2009年 第5期

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