Korean Journal of Ichthyology (한국어류학회지)

The Ichthyological Society of Korea (한국어류학회)
권호 표지

Expression of GFP Gene Driven by the Olive Flounder (Paralichthys olivaceus) hsc70 Promoter in Trangenic Medaka (Oryzias latipes)

넙치 (Paralichthys olivaceus) 열충격 유전자 hsp70 조절부위에 의한 형광단백질의 발현

  • Lee, Jeong-Ho (Genetics and Breeding Research Center, National Fisheries Research and Development Institute) ;
  • Kim, Jong-Hyun (Genetics and Breeding Research Center, National Fisheries Research and Development Institute) ;
  • Noh, Jae Koo (Genetics and Breeding Research Center, National Fisheries Research and Development Institute) ;
  • Kim, Hyun Chul (Genetics and Breeding Research Center, National Fisheries Research and Development Institute) ;
  • Kim, Woo-Jin (Biotechnology Research Institute, National Fisheries Research and Development Institute) ;
  • Kim, Young-Ok (Biotechnology Research Institute, National Fisheries Research and Development Institute) ;
  • Kim, Kyung-Kil (Genetics and Breeding Research Center, National Fisheries Research and Development Institute)
  • 이정호 (국립수산과학원 육종연구센터) ;
  • 김종현 (국립수산과학원 육종연구센터) ;
  • 노재구 (국립수산과학원 육종연구센터) ;
  • 김현철 (국립수산과학원 육종연구센터) ;
  • 김우진 (국립수산과학원 생명공학연구소) ;
  • 김영옥 (국립수산과학원 생명공학연구소) ;
  • 김경길 (국립수산과학원 육종연구센터)
  • Received : 2007.10.01
  • Accepted : 2007.11.22
  • Published : 2007.12.31
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Abstract

Heat shock proteins (HSPs) are a family of highly conserved proteins playing an important role in the functioning of unstressed and stressed cells. The HSP70 family, the most widely studied of the hsps, is constitutively expressed (hsc70) in unstressed cells and is also induced in response to stressors (hsp70), especially those affecting the protein machinery. The HSP/HSC70 proteins act as molecular chaperones and are crucial for protein functioning, including folding, intracellular localization, regulation, secretion, and protein degradation. Here, we report the identification and characterization of the putative amino acid sequence deduced from one cDNA clone identified as heat shock protein 70. The alignment showed that the putative sequence is 100% identical to the heat shock protein 70 cognate (HSC 70) of olive flounder. The 5'-flanking region sequence (approximately 1 kb) ahead of the hsc70 gene was cloned by genome walking and a putative core promoter region and transcription elements were identified. We characterized the promoter of the olive flounder hsc70 gene by examining the ability of 5'-upstream fragments to drive expression of green fluorescent protein (GFP) in live embryos.

열충격 단백질(hsp)은 세포의 기능에 중요한 역할을 하는 보존성이 높은 단백질중의 하나이다. 이들 중 70 kDa 열충격 단백질은 외부의 자극과 관계없이 상시적으로 합성되는 HSC70 단백질과 외부의 자극에 반응하여 합성되는 HSP70 단백질이 있다. 본 연구에서는 넙치(Paralichthys olivaceus)의 70 kDa 열충격 단백질에 대한 cDNA를 아미노산 서열로 변환시켜 분석함으로써 이 유전자가 상시적으로 발현하는 열충격 단백질인 HSC70에 대한 유전자임을 밝혔다. Hsp70 유전자의 발현 기작을 조사하기 위하여 단백질 발현을 조절하는 5' 인접부위를 분리하고 이들의 염기서열을 분석함으로써 유전자 조절부위의 중요인자와 중심 부위를 동정하였다. 또한 Hsp70 유전자의 유전자 조절부위를 이용하여 형광단백질 발현벡터를 제작한 후 메다카 수정란에 미세 주입하여 배 발생 과정의 살아있는 메다카에서 발현하는 형광 단백질(GFP)의 발현을 조사하였다.

Keywords

Acknowledgement

Supported by : National Fisheries Research and Development Institute

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