二硫苏糖醇提取纯化Cavl.2钙通道片段CT3蛋白

发布时间：2018-11-25 19:47

【摘要】：目的探讨二硫苏糖醇(DTT)有助于Pre Scission蛋白酶切除重组蛋白GST-CT3标签GST的方法。方法在大肠埃希菌BL21中转化入p GEX-6P-3/CT3重组质粒并诱导其表达,用B-PER提取纯化重组蛋白GST-CT3,在10 mmol/L DTT存在的情况下Pre Scission蛋白酶切除GST标签得到高浓度的CT3蛋白,进行SDS-PAGE电泳鉴定CT3蛋白。结果不含DTT情况下,Pre Scission蛋白酶很难切除重组蛋白GST-CT3的标签蛋白GST,而在10 mmol/L DTT存在的情况下,Pre Scission蛋白酶能够切除重组蛋白GST-CT3的标签蛋白GST,得到高浓度的CT3蛋白。结论 10 mmol/L DTT有助于Pre Scission蛋白酶切除重组蛋白GST-CT3的标签蛋白GST,得到高浓度的CT3蛋白。
[Abstract]:Objective to investigate the effect of dithiothreitol (DTT) on the removal of recombinant protein GST-CT3 label GST by Pre Scission protease. Methods the recombinant plasmid of p GEX-6P-3/CT3 was transformed into Escherichia coli BL21 and its expression was induced. The recombinant protein GST-CT3, was extracted and purified by B-PER. In the presence of 10 mmol/L DTT, Pre Scission protease excised the GST label to obtain high concentration of CT3 protein and identified CT3 protein by SDS-PAGE electrophoresis. Results it was difficult for, Pre Scission protease to remove the label protein GST, of recombinant protein GST-CT3 without DTT, but, Pre Scission protease could remove the label protein GST, of GST-CT3 in the presence of 10 mmol/L DTT. High concentration of CT3 protein was obtained. Conclusion 10 mmol/L DTT is helpful for Pre Scission protease to remove the label protein GST, of recombinant protein GST-CT3 to obtain high concentration of CT3 protein.
【作者单位】：中国医科大学药学院药物毒理学教研室;天津出入境检验检疫局工业品中心;
【基金】：国家自然科学基金(31471091,31400981)
【分类号】：R96

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