二硫蘇糖醇提取純化Cavl.2鈣通道片段CT3蛋白
發(fā)布時(shí)間：2018-11-25 19:47
【摘要】：目的探討二硫蘇糖醇(DTT)有助于Pre Scission蛋白酶切除重組蛋白GST-CT3標(biāo)簽GST的方法。方法在大腸埃希菌BL21中轉(zhuǎn)化入p GEX-6P-3/CT3重組質(zhì)粒并誘導(dǎo)其表達(dá),用B-PER提取純化重組蛋白GST-CT3,在10 mmol/L DTT存在的情況下Pre Scission蛋白酶切除GST標(biāo)簽得到高濃度的CT3蛋白,進(jìn)行SDS-PAGE電泳鑒定CT3蛋白。結(jié)果不含DTT情況下,Pre Scission蛋白酶很難切除重組蛋白GST-CT3的標(biāo)簽蛋白GST,而在10 mmol/L DTT存在的情況下,Pre Scission蛋白酶能夠切除重組蛋白GST-CT3的標(biāo)簽蛋白GST,得到高濃度的CT3蛋白。結(jié)論 10 mmol/L DTT有助于Pre Scission蛋白酶切除重組蛋白GST-CT3的標(biāo)簽蛋白GST,得到高濃度的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.
【作者單位】： 中國醫(yī)科大學(xué)藥學(xué)院藥物毒理學(xué)教研室;天津出入境檢驗(yàn)檢疫局工業(yè)品中心;
【基金】：國家自然科學(xué)基金(31471091,31400981)
【分類號(hào)】：R96
[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.
【作者單位】： 中國醫(yī)科大學(xué)藥學(xué)院藥物毒理學(xué)教研室;天津出入境檢驗(yàn)檢疫局工業(yè)品中心;
【基金】：國家自然科學(xué)基金(31471091,31400981)
【分類號(hào)】：R96
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