Reproductive and Developmental Biology

  • 제28권3호
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  • Pages.197-202
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  • 2004
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  • 1738-2432(pISSN)
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  • 2288-0151(eISSN)
한국동물번식학회 (The Korean Society of Animal Reproduction)
권호 표지

Retrovirus Vector System을 이용한 hPTH가 발현되는 돼지 세포의 구축

Retrovirus Vector-Mediated Construction of Porcine Cell Lines Expressing Human PTH

  • 정지연 (충북대학교 축산학과) ;
  • 구본철 (충북대학교 축산학과) ;
  • 김남형 (충북대학교 축산학과) ;
  • 권모선 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 박성환 (대구가톨릭대학교 의과대학 일반외과) ;
  • 김태완 (대구가톨릭대학교 의과대학 생리학교실)
  • Jeong J. Y. (Department of Animal Science, Chungbuk National University) ;
  • Koo B. C. (Department of Animal Science, Chungbuk National University) ;
  • Kim N. H. (Department of Animal Science, Chungbuk National University) ;
  • Kwon M. S. (Department of Physiology, Catholic University of Daegu School of Medicine) ;
  • Park S. H. (Department of General Surgery, Catholic University of Daegu School of Medicine) ;
  • Kim T. O. (Department of Physiology, Catholic University of Daegu School of Medicine)
  • 발행 : 2004.09.01
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초록

골다공증 치료제로 이용되고 있는 hPTH는 체내의 혈중 칼슘 농도를 조절하는 인간의 부갑상선 호르몬이다. 본 연구에서는 hPTH를 효율적으로 생산하는 돼지세포를 구축하고자 다양한 retrovirus vector를 이용하였는데 다음과 같은 결과를 관찰하였다. 1) hPTH 유전자의 전이를 확인하기 위해 RT-PCR을 수행한 결과, 형질전환된 모든 돼지세포에서 420 bp의 hPTH에 해당하는 단편을 확인할 수 있었으며, WPRE가 도입되지 않은 실험군보다 WPRE서열이 도입된 실험군에서 더 강한 밴드를 확인하였다. 2) ECLIA 측정 결과 hPTH 유전자가 도입된 모든 세포에서 hPTH가 생성되었으며, 특히 Tet-On system에서는 doxycycline을 처리한 실험군에서 hPTH의 발현이 유도되었음을 확인하였다. 또한 RT-PCR과 ECLIA을 수행하여 hPTH의 도입 여부와 단백질 생산을 비교한 결과, WPRE 서열이 hPTH 유전자의 downstream 위치에 도입된 실험군에서 가장 많은 단백질이 생산됨을 확인할 수 있었다.

Human PTH (parathyroid hormone) is known to be efficacious for curing osteoporesis. In this study, we attempted to construct genetically modified porcine cell lines that can ultimately be used for donor cells in nuclear transfer-mediated transgenesis. By using retrovirus vectors carrying tetracycline-regulatory expression system and WPRE (woodchuck hepatitis virus posttranscriptional regulatory element) sequence, we could control PTH expression with tetracycline and boost the promoter activity. Considering that low or constitutive expression of the transgene has been one of major problems that needs to be solved in transgenic animal studies, our results could be helpful in successful production of transgenic pigs as bioreactors.

키워드

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