小鼠未受精卵和早1-细胞期受精卵中MPF蛋白水平的检测  

Detection of M-phase promotingfactor in unfertilizedandat early 1-cell fertilized eggs of mouse

作　　者：于爱鸣[1] 陈菲[1] 宗志宏[1] 武迪迪[1] 于秉治[1] 

机构地区：[1]中国医科大学基础医学院生物化学与分子生物学教研室,辽宁沈阳110001

出　　处：《中国医科大学学报》2005年第2期105-106,110,共3页Journal of China Medical University

摘　　要：目的::检测小鼠未受精卵和早1-细胞期受精卵中M期促进因子(MPF)蛋白水平,探讨MPF在早1-细胞期受精卵中的变化.方法:利用超排卵及反复冻融裂解细胞的方法,对卵母细胞和受精卵细胞中MPF的催化亚基Cdc2、调节亚基周期素B进行Western印迹分析.结果:在500个MⅡ卵母细胞和受精卵细胞样品中,可以清楚看到各有一条粉紫色的特异免疫印迹带显示Cdc2,根据凝胶成像及分析系统的Gelworks应用软件分析,得到的强度的相对值分别是100%和约80%.用700个卵细胞进行观察时可以看到MⅡ卵母细胞显示有2条免疫印迹带显示细胞周期素B,其中一条为60～63 kDa,是周期素B;另一条则位于周期素B的下方,约40～45 kDa,且色调偏淡.而受精卵细胞样品在相应的部位则未能见到特异的免疫印迹带.结论:小鼠早1-细胞期受精卵(G1期)中的Cdc2蛋白水平仍保持在相对的较高水平;而细胞周期素B的量显著减少,难以检测.Objective: To detect level of M-phase promoting factor (MPF) in unfertilized and at early 1-cell fertilized eggs of mouse in order to investigate changes of amount of MPF protein. Methods:After the superovulation and collection of eggs, protein were extracted by freezing and thawing repeatedly. The level of MPF protein were detected by using SDS-polyacrylamide gel electrophoresis (PAGE) and Western blotting. Results:For Cdc2,the perfect results were obtained when using 500 eggs as one detection sample. According to analysis of software Gelwork, the comparative values of intensity obtained were 100% and about 80% in unfertilized and fertilized eggs, respectively. For cyclin B, 2 blotting bands were observed when using 700 oocytes. The cyclin B was at the position of 60 to 63 kDa. The other was at 40 to 45 kDa, which the intensity was less than that of cyclin B. No any blotting band was observed in the sample consisting of 700 fertilized eggs for cyclin B detection. Conclusion:The amount of Cdc2 protein in early 1-cell fertilized eggs of mouse retains a high level. The amount of cyclin B protein in early 1-cell fertilized eggs of mouse falls to the minimum level, so that it is difficult to be detected.

关 键 词：受精卵 CDC2 周期素B M期促进因子 

分 类 号：Q132.7[生物学—普通生物学]