Pronuclear DNA microinjection has been the most universal method in transgenic animal production but its success rate of transgenesis in mammals are extremely low. To address this long-standing problem, we used retrovirus- and lentivirus-based vectors carrying the enhanced green fluorescent protein (EGFP) gene under the control of ubiquitously active cytomegalovirus (CMV) promoter to deliver transgenes to bovine embryos. The rate of transgenesis was evaluated by counting EGFP positive blastocysts after injection of concentrated virus stock into the perivitelline space of the bovine oocytes in metaphase II. Among two different types of lentivirus vectors derived from FIV (feline immunodeficiency virus) and HIV (human immunodeficiency virus), the former scored the higher gene transfer efficiency; almost 100% of the blastocysts developed from the oocytes infected with FIV-based vector were EGFP positive. As for the vectors derived Com HIV lentivirus, the transgenesis rate of the blastocysts was reduced to 39%.

ABSTRACT   INTRODUCTION   MATERIALS AND METHODS    Nomenclature    Construction of Retrovirus vectors    Virus Production and Infection     In Vitro Maturation (IVM)    Microinjection of Retroviral Vectior into Perivitelline Space    In Vivo Fertilization (IVF) of Oocytes    RT-PCR Analysis    Westem Analysis   RESULTS    Expression of EGFP in Bovine Fetal Fibroblasts Infceted with either FLV-CGW of HLV-OGW Recombinant Virus    Expression of EGFP in Bovine Embryos Transduced with either FLV-CGW or HLV-CGW Recombinant Virus   DISCUSSION   REFERENCES

• Young Mi Kim(Sangil Youngseo College)
• Mo Sun Kwon(Department of Catholic University of Daegu School of Medicine)
• Bon Chul Koo(Department of Catholic University of Daegu School of Medicine)
• Teoan Kim(Department of Catholic University of Daegu School of Medicine)
• Heng-Cherl Yom(Department of Life Science, Hoseo University)
• Dae Hwan Ko(Sangji Youngseo College) Corresponding author