超表达Cdc20基因不影响牛卵母细胞第一极体排出

Over-expression of Cdc20 Gene Has No Effect on Bovine Oocytes First Polar Body Extrusion

As one of the co-activator of anaphase-promoting complex(APC),cell division cycle 20(CDC20)protein also functions as the target of the spindle assembly checkpoint(SAC),which is essential for the cell cycle regulation.To investigate the function of Cdc20 during the first polar body extrusion(PBE I),Cdc20 CDS was cloned and eukaryotic expression vector pCdc20-Venus was constructed.Using the linear pCdc20-Venus as template,the capped Cdc20-Venus mRNA was synthesized via T7 Mmessage Mmachine Kit(Ambion).Cdc20 over-expression was performed by microinjection of Cdc20-Venus mRNA into the cytoplasm of bovine oocytes.The results showed that Venus tagged Cdc20 dispersed around the nucleus in HeLa cells.In bovine oocytes,the fluorescence appeared in the whole cytoplasm.However,the PBE I rate in over-expressed group(48.9%) is not significant,compared to Venus mRNA injection group(50.9%) and non-injection group(51.1%).Our study demonstrated that the over-expression of Cdc20 in bovine oocytes does not affect the PBE I rate(P>0.05).