Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
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Cloning of An Intron of the Gene Coding for Porcine Acid-Labile Subunit(pALS) of the 150-kDa Insulin-like Growth Factor Complex and the 3' ntranslated Region of pALS Complementary DNA and Confirmation of pALS Gene Expression in Multiple Tissues

돼지 150-kDa Insulin-like Growth Factor Complex의 Acid-labile Subunit(ALS) 유전자의 Intron 및 ALS Complementary DNA의 3' 비해독 부위 Cloning과 생체조직에서의 ALS 유전자 발현 확인

  • Jin, E.J. (Department of Animal Resources Technology, Regional Animal Industry Research Center, Jinju National University) ;
  • Kim, I.A. (Department of Animal Resources Technology, Regional Animal Industry Research Center, Jinju National University) ;
  • Lee, C.Y. (Department of Animal Resources Technology, Regional Animal Industry Research Center, Jinju National University)
  • 진은정 (진주산업대학교 동물소재공학과. 동물생명산업 지역협력연구센터) ;
  • 김인애 (진주산업대학교 동물소재공학과. 동물생명산업 지역협력연구센터) ;
  • 이철영 (진주산업대학교 동물소재공학과. 동물생명산업 지역협력연구센터)
  • Published : 2004.08.31
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Abstract

본 연구는 목저은 다음과 같다: 1) 돼지에서 150-kDa temary insulin-like growth faetor(IGF)complex의 한 구성 요소인 acid-labile subunit(ALS) 유전자 intron의 존재 확인. cloning 및 돼지 ALS(porcine ALS; pALS) complementary DNA(cDNA)의 3' 비해독(untranslated) 부위(3' UT) 증폭. cloning, 2) intron-spanning primer pair를 이용한 reverse transcription-polymerase chain reaction(RT-PCR) 방법에 의한 돼지 조직에서의 ALS 유전자 발현 분포 확인 및 3) 돼지 hepatocyte에서의 ALS 유전자 발현 여부 확인. 돼지 genomic DNA를 template로 하여 PCR 방법으로 예상된는 intron 부위를 증폭하고 plasmid vector에 삽입하여 염기서열을 결정한 결과 타 종의 ALS 유전자에서와 같은 위치에 1,371-base pair(bp)의 pALS intron이 존재함을 확인하였다. 역시 본 연구에서 간에서 추출한 RNA를 주형으로 시작하여 3' rapid amplification of cDNA end(3' RACE) 방법으로 147-bp의 3'UT를 합성하고 그 염기성열을 결정하였다. RT-PCR 결과 간은 물론 조사된 모든 돼지의 내장기관(신장, 폐, 비장)과 자성 생식기관(난소, 난관, 자궁) 및 골격근육에서 ALS 유전자가 발현됨이 밝혀졌다. 또한 돼지 간 조직에 대한 in-situ hybridization 결과 hepatocyte에서 ALS 유전자가 발현됨이 확인되었다. 이상의 결과는 ALS가 혈중 IGF의 저정/조절체로서의 주기능 외에 모세혈관 밖에서도 미지의 기능이 있을 기능성을 시사한다.

Keywords

References

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