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GST和His标记的重组蛋白制备及吸附纳米ZnO的研究

Keywords: 谷胱甘肽,6聚组氨酸,基因重组,蛋白表达与纯化,亲和吸附

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Abstract:

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.coliBL21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Westernblot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

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