Abstract: pMSCV containing GFP and neo- gene was transfected into the cattle fetal auris fibroblast cells by lipofection or electrofusion was the donor. The maturation of bovine oocyte in vitro was receptor. The embryo cloning was constructed by the donor and receptor. Under different conditions,such as adding epidermal growth factor(EGF)to the culture medium improved the nuclear maturation and transgenic embryos in vitro culture, donor cells gene-transfection method, conducted to investigate the effect of three in vitro culture systems on the development of bovine embryos and to observe the development of bovine transgenic embryos in vitro in the optimal culture system will be studied. To examine the effects of EGF on the nuclear and cytoplasm maturation of cattle oocytes in vitro culture, the best concentration of EGF was 30 ng/mL, then the oocytes were activated by 6-dimethylaminopurine combined with ethanol, examining the cleavage rate, blastocyst development rate. The cleavage rate and blastocyst development rate adding 25 ng/mL EGF to medium group were significantly higher than those in control group; and its effect on cell proliferation after grafting. The different gene-transfection method was beneficial to transfer embryo. The rate of the integration was the blastocyst development rate of cattle oocytes significantly（P>0.05）; the highest mSOFaa+granulosa cell monolayer co-culture system. 

Key words: cattle; transgenic; donor cells gene-transfection; in vitro