Effect of Sodium Butyrate on GFP Expression Level in Transgenic PoIygonum tinctorium Cells

쪽(Polygonum tinctortium) 세포의 형질전환 및 쪽 세포에서 Sodium Butyrate가 Green Fluorescent Protein 발현에 미치는 영향

  • Park, Sung-Kil (Plant Metabolism Research Center & School of Biotechnology, Kyung Hee University) ;
  • Chung, Choong-Sik (Plant Metabolism Research Center & School of Biotechnology, Kyung Hee University) ;
  • Lee, Jong-Jin (Plant Metabolism Research Center & School of Biotechnology, Kyung Hee University) ;
  • Lee, Youn-Hyung (Plant Metabolism Research Center & School of Biotechnology, Kyung Hee University) ;
  • Chung, In-Sik (Plant Metabolism Research Center & School of Biotechnology, Kyung Hee University)
  • 박성길 (경희대학교 식물대사 연구센터) ;
  • 정충식 (경희대학교 식물대사 연구센터) ;
  • 이종진 (경희대학교 식물대사 연구센터) ;
  • 이윤형 (경희대학교 식물대사 연구센터) ;
  • 정인식 (경희대학교 식물대사 연구센터)
  • Published : 2001.11.30

Abstract

To examine the expression of foreign protein in Polygonum tinctorium cells, plasmid pCAMBIA1302 encoding Green Fluorescent Protein(GFP) was used to transform the cells and the expression was confirmed using Western blot analysis. When the effect of sodium buryrate on the formation of GFP was examined, cell growth was retarded at the addition of 10 mM and was stalled at more than 15 mM. The amount of GFP production was increased by 15% when 5 mM of sodium butyrate was added at three-days after inoculation as compared to at 0-day. Moreover, when sodium butyrate was added at three-days after inoculation, the amount of GFP was increased by 50% at the addition of 5 mM of sodium butyrate as compared to 10 mM.

쪽(Polygonum tinctortium) 세포에서 외래 단백질 발현을 검토하기 위하여 green fluorescent protein(GFP)가 내재하는 pCAMBIA1302를 형질 전환시켰으며 Western blot 분석에 의해 GFP의 발현을 확인하였다. Sodium butyrate가 GFP생성에 미치는 효과를 검토한 결과, 10 mM에서 세포성장이 지연되었으며, 15 mM 이상에서는 정지되었다. 세포 내 GFP 생성량은 세포 접종 후 3일째 5 mM sodium butyrate를 첨가하였을 때가 0일째 처리에 비해 120% 증가하였다. 또한 접종후 3일 후 5 mM의 sodium butyrate를 처리한 경우가 10 mM의 경우보다 GFP의 수율이 50% 증가하였다. 본 실험을 통하여 세포 접종 후 3일째, 5 mM의 농도로 처리한 sodium butyrate가 외래 단백질의 발현을 효과적으로 증가시키는 결과를 확인하였다.

Keywords