Abstract: To study the exogenous gene space-time expression, the state of positive cell growth and reduplication, apoptosis, survival percentage，and to optimize the method improving the transfection efficency, we used lipofectin method to transfect six fluorescent protein genes: pEGFP-C1、pEGFP-N3、pECFP-N1、pECFP-mito、pEYFP-N1 and pDsRed1-N1 into the in vitro cultured fibroblast cell of Mongolia sheep. The results showed: 24、48 and 72h after transferring, the expression efficiency of six kind of fluorescent protein genes were between 12.3％～63.8％.Multiple comparison analysis indicated significant difference in six groups. 24h after transferring, the fluorescence could be observed in cytoplasm and nucleus welldistributed except cryptomere vesicle ; 48h after transferring, EGFP、ECFP, and EYFP gene still expressed steadily. Granula could be observed in the region of cell nucleus. DsRed gene mainly expressed some granulaexpression product surrounding nuclear membrane. In optimized condition, there were no significant effect（P>0.05）on apoptosis ratio, positive cell shape, growth and reduplication state comparing with control group. The research is important to genetic mark, nuclear transplantation and transgenic animal clone etc.