Cdc14A在小鼠一细胞期受精卵G2/M期转换中作用机制的研究

细胞分裂周期14A蛋白(Cdc14A)是一种丝氨酸/苏氨酸双特异性磷酸酶,能够降低细胞周期素依赖激酶1(Cdk1)的活性,逆转依赖Cdk1(Cdc2)的磷酸化事件。在裂殖酵母，Cdc14在有丝分裂末期调节Cdc25的稳定性，以确保在胞质分裂前钝化Cdk1的活性。人的Cdc14A在G2/M过渡时，下调Cdc25B的活性以阻止提前激活Cdc2，避免细胞过早地进入有丝分裂。小鼠一细胞期受精卵是否表达Cdc14A以及在G2/M过渡期是否具有调控作用,迄今国内外未见报道。为了验证Cdc14A在小鼠一细胞期受精卵G2期阻滞中的作用，本研究以小鼠一细胞期受精卵为研究对象，通过RT-PCR和免疫印迹实验证实确实存在Cdc14A。通过过表达Cdc14A和Cdc14A的干涉实验探讨Cdc14A调控小鼠一细胞期受精卵G2/M转变的分子机制，为哺乳动物受精卵早期发育的机理研究提供实验依据。

CDC14A is a serine/threonine dual-specificity phosphatase that reduces CDK1 activity and reverses CDK1(Cyclin dependent kinases)-dependent phosphorylation events. In fission yeast, the Cdc14 regulates the stability of the Cdc25 at the end of mitosis to ensure Cdk1 inactivation before cytokinesis. Human Cdc14A can downregulates Cdc25B activity at the G2/M transition to prevent premature activation of Cdc2 and untimely entry into mitosis. Little is know about whether exist Cdc14A phosphatase and whether Cdc14A play a regulatory role in the G2/M transition period of mouse one-cell fertilized eggs. To test the role and function of Cdc14A phosphatase in G2 arrest of mouse one-cell fertilized eggs, our previous studies demonstrated that Cdc14A was present in mouse one-cell fertilized eggs (G1,S,G2,M phase) by RT-PCR and Western blotting. Moreover, we construct expressive vector of Cdc14A and try to explore the molecular mechanism of G2/M transition of mouse one-cell fertilized eggs by overexpression and knockdown of Cdc14A. It will be of great importance for the study of mechanisms regulating the early fertilized eggs development of human and other mammals.