【摘要】 目的纯化原核表达的Neuritin蛋白,通过观察其对PC12细胞和研究其神经生物学功能。方法采用镍离子金属螯合亲和层析法纯化Neuritin蛋白,Bradford法对纯化的Neuritin蛋白进行定量,然后加入到PC12细胞,通过观察PC12细胞突起的生长情况和细胞形态的变化,研究其促进细胞突起生长的功能。结果纯化后的Neuritin蛋白经SDS-PAGE电泳显示,获得唯一蛋白条带,即Neuritin蛋白;Bradford定量法计算出其浓度为0.45 mg/ml,纯化的Neuritin蛋白加入到PC12细胞培养液中,PC12细胞长出突起,发生类神经元样改变。结论原核表达的Neuritin蛋白得到有效纯化及复性,纯化的Neuritin蛋白能促进神经突起生长,考虑Neuritin促进突触发生和调节突触可塑性等方面具有重要作用,为实验室进一步功能研究打下了一定的基础,同时作为新的神经营养因子在治疗神经疾病及其损伤中具有良好的应用前景。更多还原

【Abstract】 Objective To purify Neuritin protein from Prokaryotic expression system of neuritin,and study its neurobiological function by observing its effects on PC12 cells. Methods Using Ni affinity chromatography to purify Neuritin protein,and Bradford method to quantify purified Neuritinprotein,then added it into the culture medium of PC12 cells. The promoting neurite outgrowth on cells and cellular morphology change was detected by observing the neurite outgrowth of PC12 cells,and the activity of promoting nerve fibers growth was detected by observing the nerve fibers growth of PC12 cells. Results The purified Neuritin protein was analyzed by SDS-PAGE,and showed only one band of protein was Neuritin protein. Its concentration was 0. 45 mg / ml through the Bradford quantitative method calculation. After adding purified Neuritin protein into PC12 cell culture medium,PC12 cells grew protrusions and neuron-like changes appeared; and which were positively correlated with the concentration. Conclusion The Neuritin protein from Prokaryotic expression system is purified successfully and it can promote the neurite outgrowth of cells,which lays a good foundation for further study on the function and mechanism of neuritin.更多还原