Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Production of Bovine Transgenic Cloned Embryos using Prourokinase-Transfected Somatic Cells: Effect of Expression Level of Reporter Gene

인간 Prourokinase가 도입된 체세포를 이용한 소 형질전환 복제란 생산: 표지유전자 발현정도에 따른 효과

  • J. K. Cho (Department of Theriogenology & Biotechnology, College of Veterinary Medicine, Seoul National University) ;
  • M.M.U. Bhuiyan (Department of Theriogenology & Biotechnology, College of Veterinary Medicine, Seoul National University) ;
  • G. Jang (Department of Theriogenology & Biotechnology, College of Veterinary Medicine, Seoul National University) ;
  • G. Jang (Department of Theriogenology & Biotechnology, College of Veterinary Medicine, Seoul National University) ;
  • Park, E. S. (School of Agriculutral Biotechnology, Seoul National University) ;
  • S. K. Kang (Department of Theriogenology & Biotechnology, College of Veterinary Medicine, Seoul National University) ;
  • Lee, B. C. (Department of Theriogenology & Biotechnology, College of Veterinary Medicine, Seoul National University) ;
  • W. S. Hwang (Department of Theriogenology & Biotechnology, College of Veterinary Medicine, Seoul National University)
  • Published : 2002.08.01
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Abstract

Human Prourokinase (proUK) offers potential as a novel agent with improved fibrin specificity and, as such, may offer advantages as an attractive alternative to urokinase that is associated with clinical benefits in patients with acute peripheral arterial occlusion. For production of transgenic cow as human proUK bioreacotor, we conducted this study to establish efficient production system for bovine transgenic embryos by somatic cell nuclear transfer (NT) using human prourokinase gene transfected donor cell. An expression plasmid for human prourokinase was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human prourokinase target gene into a pcDNA3 plasmid. Cumulus cells were used as donor cell and transfected with the expression plasmid using the Fugene 6 as a carrier. To increase the efficiency for the production of transgenic NT, development rates were compared between non-transfected and transfected cell in experiment 1, and in experiment 2, development rates were compared according to level of GFP expression in donor cells. In experiment 1, development rates of non-transgenic NT embryos were significantly higher than transgenic NT embryos (43.3 vs. 28.4％). In experiment 2, there were no significant differences in fusion rates (85.4 vs. 78.9％) and cleavage rates (78.7 vs. 84.4％) between low and high expressed cells. However, development rates to blastocyst were higher in low expressed cells (17.0 vs. 33.3％), and GFP expression rates in blastocyst were higher in high expressed cells (75.0 vs. 43.3％), significantly.

Keywords

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