en Original Article Original Article Abstract Background: Determination of transgenic embryos from non transgenic embryos sibling is an important step in producing homozygous transgenic mice. These steps need by PCR or southern blotting followed extraction of DNA, but both techniques require skill and consume time. Objective: The aim of this study was simulation of high accuracy method using novel enhanced green fluorescent protein (EGFP) gene cassette to eliminate some consume time in livestock industry to assay high quality embryos and morphological plasticity. Materials and Methods: We modified pQE-Tri systemic vector with EGFP and IRES sequence to trace out coming planning of molecular farming transgene using co-injection method. Results: The combination of these sequences successfully showed the faint and normal expression of transgene in mouse pre-implantation stage embryos. The low rate of surviving green positive embryos as compare as only medium and physical treatments could be partly from the physical damage caused by microinjection and gene integration. Furthermore, application of the enhanced GFP marker facilitated subtle detection of asymmetrical division appeared in some of transgenic embryos. Conclusion: The results of current mouse simulation model imply that an efficient production and propagation of transgenic livestock can be done by co-injection of every economical gene with this novel transgene and also it can be suitable gene cassette for numerous experiments and study of protein behaviors in living cells. Enhanced green fluorescent protein (EGFP), Transgenic animals, Embryo 151 157 http://ijrm.ir/browse.php?a_code=A-10-549-303&slc_lang=en&sid=1 Abdolhossein Rezaeian No Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Seyed Mehdi Kalantar kalantarsm@ystp.ac.ir Yes Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Safar Farajnia No Biotechnology Research Center, Iran Pasteur Institute, Tehran, Iran Mehrdad Soleimani No Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Abbas Baghi No Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Abbas Aflatoonian No Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Sirous Zeinali No Biotechnology Research Center, Iran Pasteur Institute, Tehran, Iran