种子醇溶蛋白提取及检测条件探索

Extraction and Identification of Prolamin in Different Species' Seeds

以大麦、小麦、玉米、高粱和苏丹草的种子为材料提取种子的醇溶蛋白,分析了不同提取剂及不同固液比[种子粉末样品质量(g)与提取剂的体积(mL)的比例]对种子醇溶蛋白提取效果的影响,并对SDS-PAGE检测醇溶蛋白中的不同胶浓度、厚度以及样品上样量等的影响进行了研究.结果表明,60%的正丙醇、乙二醇、异丙醇和叔丁醇分别是小麦、大麦、玉米以及高粱和苏丹草的最佳提取剂,将1∶6比例提取的种子醇溶蛋白以15μL上样,0.5mm厚度的15%分离胶电泳可以得到清晰的电泳检测图谱.

Different solvents and ratios [ （the weight of seeds＇ powder, g） / （the volume of solution, mL） ] were used to extract prolamins of barley （Hordeum vulgate）, wheat （ Triticum aestivum ）, maize （Zea may ）, kaoliang （Sorghum bicolor） and Sudan grass （Sorghum sudanense）. Effects of different solvents and ratios on extracting prolamins from different species were analyzed. Meanwhile, the effects of different factors of Sodium Dodecyl Sulfate-Polyacrylamide gel electrophoresis （SDS-PAGE） on identifying prolamin were also carried out, such as gel concentrations,gel thickness and sample volumes. The results showed that 60% of propanol, ethylene glycol,iso-propyl alcohol, tert-butyl alcohol with the ratios of 1 ： 6 was the best reagent to extracting prolamins from seeds of wheat, barley, maize, kaoliang,and sudanese grass, respectively. While using SDS-PAGE with 15 % PAGE and 0.5 mm gel thickeness, the clear and distinct patterns can be identified from 15 μL of prolamin extraction.