원문정보
초록
영어
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%.
목차
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