Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Interaction of Phenolic Compound-Specific Activator with Its Promoter using SPR-Based DNA Chip

SPR 근거 DNA 칩을 이용한 페놀 화합물 특이 CapR 조절 단백질과 촉진유전자와의 상호작용 연구

  • 박선미 (부산대학교 자연과학대학 분자생물학과) ;
  • 박후휘 (부산대학교 자연과학대학 분자생물학과) ;
  • 임운기 (부산대학교 자연과학대학 분자생물학과) ;
  • 신혜자 (동서대학교 응용공학부 환경공학전공)
  • Published : 2003.02.01
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Abstract

Aromatic compounds are of major concern among environmental pollutants due to their toxicity and persistence. To monitor aromatic compounds in a real time with a better sensitivity, a new method of SPR (surface plasmon resonance) based on DNA chip (Biacore 3000) was developed here. It is thought that CapR regulatory protein as a complex with phenol, could bind to their corresponding promoter, Po. Biotinylated DNA oligomers for the promoter was synthesized by PCR and coupled onto streptoavidin-linked CM5-chip. CapR regulatory proteins were purified after cloning their genes in pET21a (+) vector and expressing proteins. The interaction was assessed by the system where the regulatory proteins flowed with or without phenol through the cells of DNA chip. CapR regulatory protein even in the presence of phenol had no response to its promoter, Po, suggesting that other factor(s) might be required for the activation of Po promoter. The present work reveals a promising possibility of the SPR-based DNA chip in monitoring specific environmental pollutants in a real time.

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References

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