Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Effect of Transcriptional Terminator Sequences on Recombinant Protein Expression from Drosophila melanogaster S2 Cell

전사 종결 염기 서열이 Drosophila melanogaster Schneider line 2 세포에서 외래 단백질의 발현에 미치는 영향

  • Hwang, In-Sook (Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University) ;
  • Park, Jong-Hwa (Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University) ;
  • Lee, Youn-Hyung (Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University) ;
  • Yoon, Jae-Seung (Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University) ;
  • Baek, Kwang-Hee (Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University) ;
  • Chung, In-Sik (Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University)
  • 황인숙 (경희대학교 생명공학원, 식물대사연구센터) ;
  • 박종화 (경희대학교 생명공학원, 식물대사연구센터) ;
  • 이윤형 (경희대학교 생명공학원, 식물대사연구센터) ;
  • 윤재승 (경희대학교 생명공학원, 식물대사연구센터) ;
  • 백광희 (경희대학교 생명공학원, 식물대사연구센터) ;
  • 정인식 (경희대학교 생명공학원, 식물대사연구센터)
  • Published : 2001.11.30
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Abstract

Utilizing the foreign gene expression system of Drosophila melanogaster Schneider line 2(S2) cell, the degree of transient protein and mRNA expression was examined with different terminators. In the case of transient expression, the expression level of green fluorescent protein(GFP) was the highest when the transfection agent was eliminated and then cultivated for 36 to 48 hr. The terminators(SV40 p(A), SV40 small T-antigen and human gastrin 3'UTR) of the expression vector system were each cloned with endostatin; thereafter, the expression levels of the endostatin and its mRNA were compared. When the expression levels of endostatin were compared 36 hr after transfection, the SV40 p(A) terminator showed the highest expression level of endostatin and its mRNA.

Drosophila melanogaster Schneider line 2(S2)세포의 외래 단백질 발현 시스템을 이용한 외래 단백질의 한시적 발현을 검토하고, 서로 다른 terminator를 이용하였을 때의 단백질 발현 및 mRNA 발현 정도를 검토하였다. 한시적 발현의 경우 transfection agent를 제거하고 36-48시간 동안 배양한 경우, 가장 높은 green fluorescent protein(GEP)의 발현을 보였다. SV4O p(A), SV4O small T-antigen, 인간 gastrin 3'UTR을 terminator로 지니는 발현 벡터시스템에 각각 endostatin유전자를 cloning시킨 뒤 재조합 endostatin의 mRNA의 발현 정도를 비교하였다. 한시적 발현을 시킨 뒤 36시간 후 endostatin의 발현 정도를 비교해 본 결과 SV40 p(A)를 terminator로 사용했을 때 mRNA및 단백질의 발현이 가장 높았다.

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