Journal of Veterinary Clinics (한국임상수의학회지)

The Korean Society of Veterinary Clinics (한국임상수의학회)
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Differential Gene Expression in the Bovine Transgenic Nuclear Trasnsfer Embryos

소 형질전환 복제란의 유전자 이상발현 규명

  • Cho, Jong-Ki (College of Veterinary Medicine, Chungnam National University) ;
  • Song, Bong-Seok (Center of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology) ;
  • Yong, Hwan-Yul (Craniomaxillofacial Life Science BK 21, Dental Research Institute, School of Dentistry, Seoul National University) ;
  • Lee, Doo-Soo (College of Veterinary Medicine, Chungnam National University) ;
  • Koo, Deok-Bon (Center of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Kyung-Kwang (Center of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology) ;
  • Shin, Sang-Tae (College of Veterinary Medicine, Chungnam National University)
  • 조종기 (충남대학교 수의과대학) ;
  • 송봉석 (한국생명공학연구원 동물발생공학연구실) ;
  • 용환율 (서울대학교 치과대학 치학연구소 BK21 치의학생명과학사업단) ;
  • 이두수 (충남대학교 수의과대학) ;
  • 구덕본 (한국생명공학연구원 동물발생공학연구실) ;
  • 이경광 (한국생명공학연구원 동물발생공학연구실) ;
  • 신상태 (충남대학교 수의과대학)
  • Published : 2007.09.30
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Abstract

The detrimental effects of gene transfection on embryo development and the molecular mechanism behind the differential expression of genes related to early embryo development were assessed in the production of transgenic cow embryos through somatic cell nuclear transfer (NT). Parthenogenetic, IVF, and transgenic NT embryos derived from ${\alpha}_1$-antitrypsin transfected ear fibroblast cells was produced. To investigate the molecular mechanism behind lower developmental competence of transgenic NT embryos, the differential mRNA expression of three genes ($IFN-{\tau}$, Oct4, Fgf4) in the 3 types of embryo (Parthenogenetic, IVF, transgenic NT) was examined. RNA was extracted from ten blastocysts derived from 3 types of embryos and reverse-transcripted for synthesis of the first cDNA. The quantification of 3 gene transcripts ($IFN-{\tau}$, Oct4, and Fgf4) was carried out in three replicate by quantitative real-time reverse transcriptase PCR. Expression level of $IFN-{\tau}$ mRNA was significantly higher in transgenic NT embryos than parthenogenetic and IVF embryos (P<0.05). However, expression level of Oct4 and Fgf4 of transgenic NT embryos was significantly lower than IVF embryos (P<0.05). Altered levels of these three mRNA transcripts may explain some of the embryonic/fetal/neonatal abnormalities observed in offspring from transgenic NT embryos.

체세포 핵이식을 통한 형질전환 소를 생산 시 초기 수정란 발육 시 발생하는 주요 유전자의 이상 발현을 규명을 목적으로 본 연구를 수행하였다. 형질전환 복제수정란의 낮은 발육능의 원인을 규명하기 위해 하기 위하여 단위 발생란, 체외수정란 및 형질전환 복제수정란에서 초기 배 발육에 중요한 유전자인 IFN-t, Oct4 및 Fgf4유전자의 발현량을 비교 분석하였다. RNA는 각각 10개 수정란에서 추출한 후 reverse-transcription하여 first cDNA를 합성하고 이를 가지고 실시간 중합효수 연쇄반응을 실시하였다. IFN-t유전자의 발현은 형질전환 복제란에서 유의적으로 높게 나왔다 (P<0.05).그러나 Oct4 및 Fgf4 유전자의 경우 형질전환 복제란에서 체외수정란에 비해 유의적으로 낮게 나옴을 확인하였다. 이상의 결과로, 형질전환 복제수정란의 낮은 발육능이 원인이 발육에 중요한 유전자의 이상 발현에 기인된다고 사료된다.

Keywords

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