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Development of Transgenic NT Embryos Using Bovine Fetal Fibroblasts Transfected with hFSH Gene  

Yang B.C. (National Livestock Research Institute, RDA)
Im G.S. (National Livestock Research Institute, RDA)
Kim D.H. (National Livestock Research Institute, RDA)
Min K.S. (Animal Biotechnology, Graduate School of Bio. and Information Technology, Hankyong National University)
Yoon D.H. (National Livestock Research Institute, RDA)
Park H.S. (National Livestock Research Institute, RDA)
Kim S.W. (National Livestock Research Institute, RDA)
Hwang I.S. (National Livestock Research Institute, RDA)
Seo J.S. (National Livestock Research Institute, RDA)
Seong H.H. (National Livestock Research Institute, RDA)
Yang B.S. (National Livestock Research Institute, RDA)
Publication Information
Journal of Embryo Transfer / v.21, no.1, 2006 , pp. 13-20 More about this Journal
Abstract
The purpose of this study was to develope the transgenic cattle expressing hFSH into the urine using the nuclear transfer. To produce the interest gene in urine, the specific vector was ligated with hFSH gene undo. maUII promoter. The fetal fibroblast cells (KbFF) were isolated from a 45-day male fetus. The hFSH gene was co-transfected with pcDNA3 (neo) vector to KbFF cells by electroporation. The gene-transfected cells were cultured with G-418 selection medium for 2 weeks. Selected colonies were confirmed by PCR. For nuclear transfer, enucleated bovine oocytes were transferred with hFSH transfected or nontransfected fetal fibroblasts. The cleavage and blastocyst formation rates were significantly lower (p<0.05) in cloned embryos transfected with hFSH gene (68.7% and 15.7%) than in those non-transfected (67.6% and 24.5 %), respectively. Apoptosis analysis showed no difference between hFSH transfected and non-transfected blastocysts (p>0.05). The blastocysts were transfected to 77 (control 24, hFSH 53) recipient cows. Two calves were born (1.9%) following transfer with NT embryos transfected with hFSH gene, but they were confirmed not to be transgenic calves. This result shows that the hFSH colonies were mixed with transfected and non transfected cells. Further research will be needed for selection and establishment of gene transfected cells.
Keywords
electroporation; hFSH; Fetal fibroblast; nuclear transfer;
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