[目的]获得绵羊卵母细胞蛋白质组表达图谱.[方法]采用直接裂解法制备卵母细胞蛋白样品,采用Bradford法和Ramagli法测定绵羊卵母细胞蛋白质样品的浓度,并进行双向凝胶电泳,运用PDQuest8.0对2-DE图谱进行初步分析.[结果]Ramagli法能更准确地测定绵羊卵母细胞蛋白质样品的浓度,从而优化样品的上样量,得到更高质量的2-DE图谱;Bradford法测得的绵羊卵母细胞蛋白质浓度偏高.[结论]700个卵母细胞的上样量比较合理,可用于构建卵母细胞蛋白质组2-DE图谱.%[Objective] To obtain ovine oocytes 2-dimensional gel electrophoresis (2-DE) maps. [Method] Ovine oocytes were solubilized in lysis buffer,and protein concentrations were measured using the method of Bradford and Ramagli. Then the samples were subjected to two-dimensional polyacrylamide gel electrophoresis. The 2-DE maps of ovine oocytes were analyzed through PDQuest8. 0. [ Result ] The Ramagli method could reflect concentration of oocyte protein more actually and could optimize the quantity of samples to get a 2-DE map with higher quality. The protein concentration measured by the Bradford method was higher than its actual value. [ Conclusion] Each 2-DE map loading 700 oocytes is more reasonable.
展开▼
