Cloning of Acetate Kinase Gene from the Copepod Paracyclopina nana and its Expression in Escherichia coli

요각류 Paracyclopina nana Acetate Kinase의 클로닝 및 대장균에서의 발현

  • Jung Sang-Oun (Department of Molecular and Environmental Bioscience, Graduate School, Hanyang University) ;
  • Seo Jung Soo (Department of Molecular and Environmental Bioscience, Graduate School, Hanyang University) ;
  • Lee Young-Mi (Department of Molecular and Environmental Bioscience, Graduate School, Hanyang University) ;
  • Park Tae-Jin (Department of Molecular and Environmental Bioscience, Graduate School, Hanyang University) ;
  • Kim Il-Chan (Polar BioCenter, Korea Polar Research Institute, Korea Ocean Research & Development Institute) ;
  • Park Heum Gi (Division of Marine Resource Development, College of Life Sciences, Kangnung National University) ;
  • Lee Jae-Seong (Department of Molecular and Environmental Bioscience, Graduate School, Hanyang University)
  • 정상운 (한양대학교 대학원 분자생명환경과학과) ;
  • 서정수 (한양대학교 대학원 분자생명환경과학과) ;
  • 이영미 (한양대학교 대학원 분자생명환경과학과) ;
  • 박태진 (한양대학교 대학원 분자생명환경과학과) ;
  • 김일찬 (한국해양연구원 극지연구소) ;
  • 박흠기 (강릉대학교 생명과학대학 해양생명공학) ;
  • 이재성 (한양대학교 대학원 분자생명환경과학과)
  • Published : 2005.09.01

Abstract

The acetate kinase gene from the copepod Paracyclopina nana was cloned. The open reading frame (ORF) was 1,200 bp, and poly(A) signal sequence was located in the end of the ORF. After the molecular phylogenetic analysis of P nana acetate kinase gene, it was revealed that it formed the same branch with that of Aspergillus. Also P. nana acetate kinase showed the difference with those of other prokaryotic microorganisms but showed the same clade with those of fungi. We also confirmed that the recombinant protein of P. nana acetate kinase made approximately 50 kDa after expression of recombinant gene construct in E. coli. This may be useful to compare this protein to those of other organisms in biochemical characteristics.

요각류 Paracyclopina nana Acetate Kinase를 클로닝하였다. 전체 open reading frame은 1,200 bp이었으며, poly(A) signal sequence가 ORF에 내재되어 있었다. 분자계통학적 분석결과 P. nana acetate kinase 유전자는 진핵생물계 곰팡이류인 Aspegillus와 같은 branch를 형성하였고, P. nana acetate kinase가 다른 원핵미생물들의 acetate kinase와는 구별되며 fungi와 같은 branch에 존재하는 것을 확인하였다. 또한, E. coli를 이용하여 원핵세포 발현벡터를 이용한 단백질 발현 유도를 통하여 P. nana acetate kinase 단백질 분자량이 약 50 kDa에 이르는 것을 확인하였다. 이 자료는 본 요각류와 다른 생물의 acetate kinase 단백질의 생화학적 특성비교에 유용하게 쓰이리라 사료된다.

Keywords

References

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