关键词: 蛋白激酶CK2β；siRNA；M期促进因子；小鼠受精卵

Abstract:

Abstract:Objective To study the influence of the regulatory subunit of protein kinase CK2β in the early development of fertilized mouse eggs by the means of specific siRNA and to clarify the mechanism of regulation of early development of fertilized mouse eggs.Methods One-cell stage fertilized mouse eggs were collected after superovulation,and microinjected with TE buffer,scramble siRNA or CK2β siRNA,then the effect of siRNA  was examined by RT-PCR and Western blotting.After RNA interference,the morphological changes and cleavage rate of mouse embryos were observed under dissecting microscope,and the M-phase promoting factor（MPF）activity of fertilized mouse eggs were detected by autoradiography.Results Compared with TE buffer and scramble siRNA groups,the expression levels of CK2β mRNA and protein in CK2β siRNA group were significantly reduced（P<0.01）; the activity of MPF in CK2β siRNA group reached its peak value at 28.5 h after human chorionic gonadotropin（hCG) injection,about 30 min earlier than TE buffer and scramble siRNA groups.The embryos treated with CK2β siRNA entered the M phase at 28.5 h after hCG injection,and the cleavage rate was as high as 89.5% at 32.0h,markedly higher than those in TE buffer and scramble siRNA groups (P<0.01).Conclusion CK2β siRNA may promote the early development of fertilized mouse eggs via activating MPF activity ahead of time.

Key words: CK2β；siRNA；M-phase promotingfactor；fertilized mouse eggs