| GST和His标记的重组蛋白制备及吸附纳米ZnO的研究 |
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| 引用本文： | 宋磊,陈劲春.GST和His标记的重组蛋白制备及吸附纳米ZnO的研究[J].北京理工大学学报,2011,31(2):240-243. |
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| 作者姓名： | 宋磊 陈劲春 |
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| 作者单位： | 北京化工大学,生命科学与技术学院,北京,100029 |
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| 摘 要： | 以pET-28a(+)载体为模板人工设计引物,通过PCR反应扩增带有质粒上游6聚组氨酸(6×Histidine)序列的目的基因片段,克隆到原核表达载体pGEX-4T-3中,筛选及鉴定阳性重组子pGEX-His,转化E.coli BL21细胞后通过IPTG诱导表达.由谷胱甘肽(GST)和6聚组氨酸标记的重组蛋白GST-His得到表达,利用Ni-NTA系统进行纯化.通过SDS-PAGE电泳和Western blot分析鉴定,由LC-MS/MS质谱分析确定所得蛋白为目标产物.通过蛋白结合纳米无机材料的亲和吸附实验结果证实,重组的新蛋白表现出对纳米ZnO特异吸附的生物活性.
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| 关 键 词： | 谷胱甘肽 6聚组氨酸 基因重组 蛋白表达与纯化 亲和吸附 |
| 收稿时间： | 2010/7/25 0:00:00 |
Preparation of Recombinant Protein with GST and His Tag and Its ZnO-Binding Quality |
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| SONG Lei and CHEN Jin-chun.Preparation of Recombinant Protein with GST and His Tag and Its ZnO-Binding Quality[J].Journal of Beijing Institute of Technology(Natural Science Edition),2011,31(2):240-243. |
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| Authors: | SONG Lei and CHEN Jin-chun |
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| Institution: | SONG Lei,CHEN Jin-chun(College of Life Science and Technology,Beijing University of Chemical Technology,Beijing 100029,China) |
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| Abstract: | The plasmid pET-28a(+) was used as templates to PCR-amplify objective fragment containing the upstream 6×Histidine sequence with designed primers.PCR products were ligated into a prokaryotic expression vector pGEX-4T-3.Positive E.coli clones containing recombinant plasmid pGEX-His were selected and identified,followed by transformation into E.coli BL21 cells and induction with IPTG.The recombinant protein GST-His was expressed with GST and His tag,purified with Ni-NTA system,identified by SDS-PAGE electrophoresis and Western blot analysis and confirmed by LC-MS/MS analysis.Affinity adsorption test on the protein’s binding with nanoparticles demonstrated that the fusion protein had a specific adsorption with ZnO nanoparticles. |
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| Keywords: | glutathione-S-transferase 6×Histidine genetic recombination protein expression and purification affinity adsorption |
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