Embryos that were produced by parthenogenesis were cultured to morula, then each cell of morulas was transferred into enucleate MⅡ oocytes, as called parthenogenetic morula nuclear transplantation embryos here. NIH3T3 cell nucleus were also transferred into enucleate MⅡ oocytes. In order to prove up the relationship between nuclear reprogram and DNA methylation during the process of nuclear transplantation, the two cloned pre-implantation embryos together with in vitro fertilization (IVF) and parthenogenetic embryos were subjected to immunocytochemistry with antibody to DNA methylation (5'-MeC). In order to research the regulation of oocyte cytoplasm to the relative expression level of imprint genes, the relative expression level of U2afbp-rs gene which is maternally imprint and non-imprint eIF-4C gene in pre-implantation IVF, parthenogenetic and parthenogenetic morula nuclear transplantation embryos by real-time PCR were detected.The results show that neither the genomic DNA of NIH3T3 cloned embryos nor parthenogenetic morula nuclear transplantation embryos were demethylated actively. Although both the relative expression level of U2afbp-rs gene and eIF-4C gene in parthenogenetic morula nuclear transplantation embryos were lower than control parthenogenetic embryos, the expression of the two genes were similar to the control parthenogenesis group. These results suggest that the imprinting gene expression of donor nuclear are regulated by oocyte cytoplasm.