Isolation and Characterization of Nucleoside Diphosphate Kinase 1 of Codonopsis lanceolata

더덕에서 Nucleoside Diphosphate Kinase 1 분리 및 분석

  • 김종학 (㈜바이오피아) ;
  • 양덕춘 (경희대학교 생명과학대학 및 한방재료가공센타)
  • Published : 2003.10.01

Abstract

The NDK1 is an ubiquitous enzyme that transfer phosphate groups from triphosphate nucleoside diphosphates(NDPs) in eukaryotes and prokaryotes. We isolated and characterized a cDNA encoding a nucleoside diphosphate kinase 1(CNDK 1) in Codonopsis lanceolata. The CNDK 1 is 444bp long and open reading frame of 447bp with a deduced amino acid of 148 residue. The CNDK 1 has an ATP binding site in 12­16 residue and phosphohistidine intermediate in 115 residue of amino acid sequence. Although several NDK 1 genes have been cloned in plants, but little is known about the functional significance of this enzyme during plant growth and development. The CNDK 1 shows the identities to Arabidopsis thaliana (71%), Oryza sativa(75%), Glycine max (79%), Brassica rapa (77%), Mesembryanthemum crystallinum (85 %), Spinacia oleracea (83%), Pisum sativum (82%). The CNDK 1 of C. laceolata have a closer relationship of Glycine max and Pisum sativum at the phylogenic analysis.

더덕의 재배는 수익성이 높고 재배면적도 증가하지만 수요를 만족시킬 만큼 공급이 따르지 못하고 있다. 또한 재 배 상의 어려운점은 병충해, 기계수확에 의한 대규모 재배를 더욱더 곤란하게 하고 있는 실정이다. 이러한 문제점 및 환경적 스트레스에 저항하여 자랄 수 있는 식물체를 얻기 위해 더덕의 cDNA를 분석하여 스트레스 관한 유전자 Nucleoside diphosphates kinase 1(NDK 1)을 분리하여 분석하여 148개의 아미노산 서열과 다른 식물체들의 NDK 1과 높은 유사성을 가진다는 것을 알았고, 더덕의 각 조직에서 나타나는 ClNDK1의 발현을 알아보기 위해 캘러스, 잎, 줄기, 뿌리 조직의 전체 RNA를 추출하여 cDNA를 합성하고 PCR을 수행하였다 RT­PCR 분석 결과, ClNDK1은 조직 특이성 없이 캘러스, 잎, 줄기, 뿌리 조직에 대해서 모두 발현이 되었으며, 발현량, 역시 큰 차이 없이 모든 조직에서 동일하게 발현되었다. NDKs 는 환경 스트레스에 저항성을 가진다고 알려져 있지만 NDK 1 대한 연구는 아직까지 부족한 상태이다. 우리는 더덕에서 분리한 ClNDK1의 스트레스 저항성에 대해서 지속적으로 연구를 수행 할 것이다.

Keywords

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