Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Characterization of the Promoter Controling the Stage-Specific Gene Expression of Bombyx mori

누에를 이용한 시기 특이적 발현 조절 유전자 promoter 개발

  • Park, Seung-Won (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA) ;
  • Choi, Gwang-Ho (Research Policy Planning Division, Rural Development Administration) ;
  • Goo, Tae-Won (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA) ;
  • Kim, Seong-Ryul (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA) ;
  • Kang, Seok-Woo (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA)
  • 박승원 (농촌진흥청 국립농업과학원 농업생물부 잠사양봉소재과) ;
  • 최광호 (농촌진흥청 연구정책국 연구정책과) ;
  • 구태원 (농촌진흥청 국립농업과학원 농업생물부 잠사양봉소재과) ;
  • 김성렬 (농촌진흥청 국립농업과학원 농업생물부 잠사양봉소재과) ;
  • 강석우 (농촌진흥청 국립농업과학원 농업생물부 잠사양봉소재과)
  • Received : 2011.08.19
  • Accepted : 2011.09.16
  • Published : 2011.10.31
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Abstract

We characterized embryo early gene (EEG)-704 promoter of the silkworm Bombyx mori, which is specifically regulated in the development stages. To determine core promoter region, 10 different partial mutant clones were tested by luciferase assay in Sf9 cells. About 1.5 kb promoter shows higher luciferase activity than constitutive promoter (BmA3). Interestingly, EEG-704 shares the same DNA sequences with BmHsp20.8 by the result of BLAST analysis; its expression is also increased under heat shock condition. Development of such promoter inducible, directly or indirectly in the developmental-stage, is very useful in making recombinant proteins in transgenic silkworms.

본 연구에서는 누에의 초기 배아시기에 유전자 발현 조절이 가능한 EEG-704 promoter를 개발하고자 하였다. Promoter의 핵심 영역을 결정하기 위하여, 10개의 서로 다른 partial mutant clone들을 만들고 이를 Sf9 곤충세포주에 도입하여 luciferase assay 방법을 사용하여 각각의 clone의 활성을 분석하였다. Constitutive promoter인 BmA3 promoter에 의한 활성과 비교하였을 때, 약 1.5 kb의 promoter 염기서열을 포함하는 clone이 가장 높은 luciferase 발현율을 나타내었다. 특히 EEG-704 유전자의 경우 BLAST를 이용한 유전자 비교 분석의 결과 누에의 열충격 단백질20.8 (BmHsp20.8) 과 동일한 것으로 밝혀졌으며, 정상 온도조건과 비교하였을 때 열충격을 가한 조건하에서 발현율이 증가하는 현상을 나타내었다. 특이적으로 발생단계에서 직 간접적으로 발현 조절이 가능한 이러한 promoter는 여러 유용 재조합 단백질 생산을 위한 형질전환 누에 개발 시 매우 유용할 것으로 생각된다.

Keywords

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