| 对虾白斑病毒核糖核苷酸还原酶基因原核表达载体的构建及蛋白表达、纯化 |
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| 引用本文： | 吴文林,杨培峰,陈宇,陈曦.对虾白斑病毒核糖核苷酸还原酶基因原核表达载体的构建及蛋白表达、纯化[J].泉州师范学院学报,2012,30(2):63-66. |
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| 作者姓名： | 吴文林 杨培峰 陈宇 陈曦 |
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| 作者单位： | 泉州师范学院化学与生命科学学院,福建泉州,362000 |
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| 基金项目： | 国家自然科学基金资助项目(31072236);福建省教育厅项目(JK2009043) |
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| 摘 要： | 采用PCR方法扩增对虾白斑病毒核糖核苷酸还原酶基因,插入到pGEX-4T-2表达载体上构建出带有目的基因的重组质粒pGEX-4T-2RR,然后将重组质粒转化大肠杆菌.转化菌经IPTG诱导后大量表达重组蛋白,通过降低诱导温度获得可溶性表达的重组蛋白,采用Glutathione Sepharose 4B亲和层析对重组蛋白进行纯化,获得了高纯度的目的蛋白.
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| 关 键 词： | 对虾白斑综合症病毒 核糖核苷酸还原酶 表达 纯化 |
Cloning and Expression of White Spot Syndrome Virus(WSSV) Ribonucleotide Reductase |
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| WU Wen-lin,YANG Pei-feng,CHEN Yu,CHEN Xi.Cloning and Expression of White Spot Syndrome Virus(WSSV) Ribonucleotide Reductase[J].Journal of Quanzhou Normal College,2012,30(2):63-66. |
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| Authors: | WU Wen-lin YANG Pei-feng CHEN Yu CHEN Xi |
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| Institution: | (School of Chemistry and Life Science,Quanzhou Normal University,Fujian 362000,China) |
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| Abstract: | In this study,ribonucleotide reductase gene of WSSV was amplified by PCR,digested by Sma I and Not I,and inserted into the expression vector pGEX-4T-2 to construct recombinant plasmid.Colonies with recombinant plasmid were picked for PCR and culture identified by enzyme digestion.The results show that ribonucleotide reductase gene is successfully inserted into the pGEX-4T-2 expression vector.After induced with IPTG at 18 ℃,the fusion protein is purified with Glutathione Sepharose 4B. |
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| Keywords: | white spot syndrome virus ribonucleotide reductase expression purification |
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