苏丹草种子醇溶蛋白提取和电泳条件优化

以苏丹草(Sorghum sudanense)种子为材料,以蛋白质提取率和电泳图谱效果为标准,分析不同提取剂、种子粉末质量(g)与提取剂体积(mL)的比例、还原剂二硫苏糖醇(dithiothreito,DTT)的浓度对种子醇溶蛋白提取效果的影响,同时研究上样量、不同胶浓度对SDS-PAGE检测蛋白的影响.结果表明,苏丹草的最佳提取剂为含有0.2%DTT的70%正丙醇,种子粉末质量与提取剂体积的最佳比例为1∶8.在上样量适当的情况下,12%分离胶电泳可以得到清晰的电泳图谱.以苏丹草种子为材料所建立的醇溶蛋白提取和电泳条件,也可应用于其他几种常见的禾本科种子醇溶蛋白的分析,并可得到较好的分辨率.

Optimization of Extraction and Electrophoresis of Prolamin from Sorghum sudanense seeds

This paper studies the effects of different extraction solution,ratio of sample-weight to extraction-solution and concentrations of reduction reagent dithiothreito(DTT) on extraction efficiency and quality of electrophoresis of prolamin from seeds of Sorghum sudanense.Contribution of the gel concentration and loading dosage to the results of SDS-PAGE were analyzed by evaluating the electrophoresis.The results show that 70% propylalcohol supplemented with 0.2% DTT is the best extraction solution and the ratio of sample-weight to extraction-solution 1∶8(g/mL) can give optimal results.SDS-PAGE of 12% produces the best clearance for electrophoresis under a loading appropriate sample.Established condition based on prolamin from seeds of Sorghum sudanense can be applied to isolation and electrophoresis for prolamins from several other gramineae seeds with good results.