| 酿酒酵母金属硫蛋白的表达、纯化及活性检测 |
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| 引用本文： | 梁晓峰,龚映雪,肖文娟,胡佳,郑阳阳,张小洪,王峻梅,刘泽寰.酿酒酵母金属硫蛋白的表达、纯化及活性检测[J].兰州大学学报(自然科学版),2011(3):58-62,67. |
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| 作者姓名： | 梁晓峰 龚映雪 肖文娟 胡佳 郑阳阳 张小洪 王峻梅 刘泽寰 |
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| 作者单位： | 暨南大学生命与健康工程研究院分子生物研究中心; |
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| 基金项目： | 珠海市科技攻关项目(PC20061065) |
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| 摘 要： | 通过PCR从酿酒酵母基因组DNA上扩增得到酿酒酵母基因CUP1编码的金属硫蛋白序列.将其编码序列克隆到质粒pTWIN1构建重组表达质粒pTWIN1-MT,转化大肠杆菌感受态细胞ER2566.重组融合蛋白CBD-intein1-MT在IPTG的诱导下得到表达,经SDS-PAGE和蛋白质印迹鉴定.用重组菌分别进行铜离子耐受...
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| 关 键 词： | 金属硫蛋白 酿酒酵母 原核表达 活性检测 |
Expression,purification and bioactivity analysis of Saccharomyces cerevisiae metallothionein |
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| LIANG Xiao-feng,GONG Ying-xue,XIAO Wen-juan,HU Jia,ZHENG Yang-yang,ZHANG Xiao-hong,WANG Jun-mei,LIU Ze-huan Research Center for Molecular Biology.Expression,purification and bioactivity analysis of Saccharomyces cerevisiae metallothionein[J].Journal of Lanzhou University(Natural Science),2011(3):58-62,67. |
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| Authors: | LIANG Xiao-feng GONG Ying-xue XIAO Wen-juan HU Jia ZHENG Yang-yang ZHANG Xiao-hong WANG Jun-mei LIU Ze-huan Research Center for Molecular Biology |
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| Institution: | LIANG Xiao-feng,GONG Ying-xue,XIAO Wen-juan,HU Jia,ZHENG Yang-yang,ZHANG Xiao-hong,WANG Jun-mei,LIU Ze-huan Research Center for Molecular Biology,Institutes of Life and Health Engineering,Jinan University,Guangzhou 510632,China |
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| Abstract: | The gene fragment of metallothionein coded by gene CUP1 was amplified using PCR from Saccharomyces cerevisiae genome,and was cloned into plasmid pTWIN1 to construct the recombinant plasmid pTWIN1-CBD-MT.It was then transformed into the competent E.coli ER2566 cells.The recombinant fusion protein CBD-intein1-MT was induced by IPTG and identified by SDS-PAGE and the western blot.The results of resistance and accumulation of copper ions using recombinant strain showed that the expression of metallothionein inc... |
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| Keywords: | metallothionein Saccharomyces cerevisiae prokaryotic expression bioactivity analysis |
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