The two-dimensional electrophoresis(2-DE) technique for Porcine(Sus scrofa) oocyte proteome analysis was established.The components of protein lysis solution,sample treatment,and 2-DE procedure were optimized.A profile of proteins extracted from Porcine oocytes was obtained clearly.The protein expression profiles of immature and in vitro matured Porcine oocytes were compared. The ImageMaster 2-DE software was used to characterize different proteins.About 800 protein spots were identified,of which 22 proteins were up-regulated and 13 proteins were down-regulated during oocyte maturation in vitro.It proves that proteomics technique base on two-dimensional electrophoresis can be used in further studies on differential protein expressions in Porcine oocyte development.