在体细胞有丝分裂进程中，Polo-like激酶１(Plk1)对G2/M期转化，启动有丝分裂等具有重要的调控作用，但该激酶是否也参与猪克隆（SCNT）胚胎早期有丝分裂的调控目前仍不明确。本研究拟以猪体受精(IVF)胚胎为参照，采用免疫印迹结合免疫荧光等方法检测Plk1在猪SCNT胚胎早期有丝分裂进程中的动态表达与亚细胞定位；同时利用激光共聚焦成像技术观察Plk1动态表达与微管、微丝等细胞骨架蛋白分布的时空相关性；在此基础上，通过RNAi与Volasertib抑制激酶活性等方法研究Plk1表达与激酶活力对猪SCNT胚胎有丝分裂进程的影响；最后通过AZD6244特异性抑制试验进一步分析Plk1是否通过MEK-ERK信号通路参与调控猪SCNT胚胎早期有丝分裂进程。本研究旨在揭示Plk1在猪SCNT胚胎早期有丝分裂进程中的功能及其可能的分子机制，为深入研究克隆胚胎早期发育机制提供实验参考。

Polo-like kinase 1 (Plk1) play an important role in regulation of G2/M phase transformation, and starting mitotic events in somatic mitosis process, however, its roles in porcine cloned embryonic mitosis is still unclear. IVF embryos will be used as the control, western blotting and immunofluorescent techniques will be performed to analyze the expression and subcellular localization of Plk1 in porcine cloned embryos early mitotic progression. Temporal and spatial relationship between Plk1 and cytoskeletal proteins will also be determined by confocal microscopy. To investigate the significance function of Plk1 in porcine cloned embryonic mitosis process, the expression of Plk1 will be knocked down by microinjection of Plk1 shRNA plasmid or the kinase activity of Plk1 will be inhibition by Volasertib．We will detect the expression of Plk1 and its kinase activity changes after inhibition of MEK-ERK signaling．The aim of this study is to determine the effects of Plk1 on the mitosis of porcine SCNT embryos, so as to establish a basis for the further research on the development mechanism of porcine cloned embryos.