人表皮生长因子融合蛋白在大肠杆菌表达的研究

Studies on the expression of fusion protein of recombinant hEGF in E.coli

在大肠杆菌中高效表达人表皮生长因子(hEGF)。根据大肠杆菌对密码的偏爱性,构建高效表达EGF融合蛋白的菌株,经离子交换层析和凝胶过滤层析两个步骤使产物得到纯化。融合蛋白表达产量为27%,EGF融合蛋白纯化产物纯度为95%以上,促3T3细胞增殖的活性测定为ED50为4.2ng/mL。在pET 3c:BL21(DE3)大肠杆菌表达系统中,以融合蛋白方式表达人表皮生长因子,表达效率高,纯化路线简单,产量较高,适合产业化生产。

To express hEGF in E.coli. A new-designed hEGF gene was expressed in fusion protein according to the code preference of the expression system . The fusion protein was purified by DEAE-ion exchange chromatography and gel filtration. The fusion protein was expressed in a high expression level of about 27% of the total cell protein, the purity of fusion protein after purification was up to 95%, the activity of enhancement of prolification of 3T3 cells was measured by ED50 which was 4.2ng/ml. The fusion protein of hEGF was expressed in high level, its purification is simple and with high yield, the whole procedure is suitable to large-scale production.