蔗糖浓度对渗透休克法提取细胞周质重组蛋白的影响

Effect of Sucrose Concentration on Periplasm Recombination Protein Extracted by Osmotic Shock

[目的]研究不同蔗糖浓度的高渗液对渗透休克法(Osmotic Shock)提取细胞周质蛋白的影响。[方法]将已经构建好的含Fbp A基因的表达质粒转化到EC感受态细胞中,分别在20%、30%、40%和50%的蔗糖高渗液条件下用Osmotic Shock纯化蛋白,SDS-PAGE电泳检测蛋白。[结果]30%和40%的蔗糖高渗液能得到浓度更高的重组蛋白,而用20%和50%的蔗糖高渗液提取蛋白产量较低。[结论]30%和40%的蔗糖高渗液能提高Osmotic Shock法提取细胞周质重组蛋白产量。

[ Objective] To study the effect of hypertonic solutions with different sucrose concentrations on-periplasm protein extracted by Os- motic Shock method. [ Method] The constructed vector with FbpA gene was transformed to EC competent cell. The recombination proteins were extracted by Osmotic Shock method with 20%, 30% , 40% and 50% sucrose respectively. The protein was detected by SDS-PAGE electropho- resis. [ Result] The result showed that using 30% and 40% sucrose hypertonic solutions could receive more recombination proteins, while 20% and 50% sucrose solutions received fewer protein. [ Conclusion] 30% and 40% sucrose hypertonic solutions could increase the production of periplasm recombination protein extracted by Osmotic Shock.