Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Cloning and Expression of Antifungal Protein (PR5) Genes from Hot Pepper (Capsicum annuum L.)

고추(Capsicum annuum)의 항균성 단백질(PR-5) 유전자의 클로닝과 발현 분석

  • Park, Hae-Jin (Department of Agronomy, College of Agriculture, Kyungpook National University) ;
  • Lee, Jung-Hoon (Department of Agronomy, College of Agriculture, Kyungpook National University) ;
  • Yoon, Yong-Hwi (Department of Agronomy, College of Agriculture, Kyungpook National University) ;
  • Kim, Hak-Yoon (Faculty of Environmental Studies, Keimyung University) ;
  • Shin, Dong-Hyun (Department of Agronomy, College of Agriculture, Kyungpook National University) ;
  • Lee, In-Jung (Department of Agronomy, College of Agriculture, Kyungpook National University) ;
  • Kim, Dal-Ung (Department of Agronomy, College of Agriculture, Kyungpook National University) ;
  • Kim, Kil-Ung (Department of Agronomy, College of Agriculture, Kyungpook National University)
  • 박해진 (경북대학교 농과대학 농학과) ;
  • 이정훈 (경북대학교 농과대학 농학과) ;
  • 윤용휘 (경북대학교 농과대학 농학과) ;
  • 김학윤 (계명대학교 환경학부) ;
  • 신동현 (경북대학교 농과대학 농학과) ;
  • 이인중 (경북대학교 농과대학 농학과) ;
  • 김달웅 (경북대학교 농과대학 농학과) ;
  • 김길웅 (경북대학교 농과대학 농학과)
  • Published : 2002.06.01
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Abstract

We have isolated and artificially expressed three cDNA clones of Capsicum annuum PR5 genes for elucidating the antifungal activity against Phytophthora capsici which contracted a hot pepper root rot in field condition. Three divergent PR5 proteins from hot pepper were designated as CAPR5-1 and CAPR5-2 from susceptible cultivar (Subicho) as well as CAPR5-3 from resistant cultivar (CM331) in response to P. capsici. The cDNA similarity was found over 80% of identity among the three CAPR5s, and deduced amino acid sequence was characterized that all of CAPR5s contained 16 cysteine residues which possibly had a significant role in the structural formation. The result of genomic DNA blot showed that CAPR5-1 and CAPR5-2 existed as single copy in the Subicho genome. Three recombinant CPARs in E. coli were identified by SDS-PACE, and each expressed protein was treated on the PDA medium which contained cultured pathogens. Although three CAPR5 proteins did not affected the hyphal growth of Glomerella glycines and Colletotrichum fagenarium, CAPR5-1, CAPR5-2, and CAPR5-3 showed a specific antifungal activities against P. capsici.

식물은 병원균이나 여러 가지 환경스트레스에 내하여 사기 방어기작을 가지며, 특히 PR 단백질은 병원균의 침입시에 동물의 면역반응과 유사한 생체방어반응을 나타내는 중요한 역할을 하는 것으로 알려져 있다. 본 연구에서는 고추에서 항균 특성을 나타내는 PR5 유전자를 클로닝하고 이들의 특성을 구명하였다. 고추에서 서로 다른 3종의 PR5 유전자, CAPR5-1, CAPR5-2, CAPR5-3를 클로닝하였다. 이들 유전자의 특성을 조사하고 아미노산 수준에서 유사성을 비교하여 본 결과, 서로간에는 90% 이삳의 상동성을 나타내었고 이들의 2차구조를 비교한 결과 중요한 domain은 높은 상동성을 나타내어 PR5 유전자들이 항균 특성을 나타내는데 매우 중요한 motif로 작용할 것으로 사료된다. CAPR5-1, CAPR-2, CAPR5-3 유전자들의 항균성 정도를 조사하기 위하여 이들 유전자를 대장균에서 발현시켜 단백질을 분리하여 고추 역병균인 phytophthora capsici에 처리한 결과, 균사의 성장이 억제되어 CAPR5-1, CAPR5-2, CAPR5-3 단백질들이 항균성을 지니고 있는 것으로 나타났다.

Keywords

References

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