The Korean Journal of Malacology (한국패류학회지)

The Malacological Society of Korea (한국패류학회)
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Phylogenetic Analysis based on Metallothionein Gene Sequence of an Indigenous Species Pisidium (Neopisidium) coreanum in Korea

한국 고유종 Pisidium (Neopisidium) coreanum (산골조개) 의 metallothionein 유전자를 기초로 한 분자계통 분류학적 연구

  • Baek, Moon-Ki (Department of Parasitology, Inje University College of Medicine) ;
  • Lee, Jun-Seo (Department of Parasitology, Inje University College of Medicine) ;
  • Kang, Se-Won (Department of Parasitology, Inje University College of Medicine) ;
  • Lee, Jae-Bong (Department of Parasitology, Inje University College of Medicine) ;
  • Kang, Hyun-Jung (Fisheries Resource Research Department, National Fisheries Research and Development Institute) ;
  • Jo, Yong-Hun (Department of Agricultural Biology, Chonnam National University) ;
  • Noh, Mi-Young (Department of Agricultural Biology, Chonnam National University) ;
  • Han, Yeon-Soo (Department of Agricultural Biology, Chonnam National University) ;
  • Choi, Sang-Haeng (Industrial Biotechnology & Bioenergy Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Chae, Sung-Hwa (Research Institute of GnC BIO Co., LTD.) ;
  • Park, Hong-Seog (Industrial Biotechnology & Bioenergy Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Jun-Sang (Division of Life Sciences, College of Natural Sciences, Kangwon National University) ;
  • Lee, Yong-Seok (Department of Parasitology, Inje University College of Medicine)
  • 백문기 (인제대학교 의과대학 기생충학교실) ;
  • 이준서 (인제대학교 의과대학 기생충학교실) ;
  • 강세원 (인제대학교 의과대학 기생충학교실) ;
  • 이재봉 (인제대학교 의과대학 기생충학교실) ;
  • 강현정 (국립수산과학원 자원연구과) ;
  • 조용훈 (전남대학교 농과대학 식물생명공학부) ;
  • 노미영 (전남대학교 농과대학 식물생명공학부) ;
  • 한연수 (전남대학교 농과대학 식물생명공학부) ;
  • 최상행 (한국생명공학연구원 바이오화학에너지센터) ;
  • 채성화 ((주)지앤시바이오) ;
  • 박홍석 (한국생명공학연구원 바이오화학에너지센터) ;
  • 이준상 (강원대학교 자연과학대학 생명과학부) ;
  • 이용석 (인제대학교 의과대학 기생충학교실)
  • Received : 2009.06.25
  • Accepted : 2009.08.10
  • Published : 2009.08.31
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Abstract

Pisidium (Neopisidium) coreanum is a freshwater snail and lives in spring water near mountain areas. Interestingly, this snail has been traditionally regarded as medicinal food, and thus has been used as folk remedies for healing broken bones. Recently, alpha classification on Pisidium (Neopisidium) coreanum through redescription has been conducted. However, not much attention has been made in beta classification. In this study, we performed the beta classification based on metallothionein (MT) genes found from various organisms. To this end, the complete cDNA sequences were obtained from the Expressed Sequence Tag (EST) sequencing project of Pisidium (Neopisidium) coreanum. The coding region (315 bp) encoded an amino acid sequence of 105 residues. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of Pc-MT gene indicate that Pisidium (Neopisidium) coreanum has similarity to freshwater bivalve such as Dreissena polymorpha (zebra mussel), Unio tumidus (swollen river mussel) and Crassostrea ariakensis (suminoe oyster).

Pisidium (Neopisidium) coreanum의 metallothionein 유전자는 염기서열 315개로 이루어져있으며 105개의 아미노산으로 이루어져 있었다. 연체동물의 metallothionein 서열의 공식[C-x-C-x(3)-C-T-G-x(3)-C-x-C-x(3)-C-x-C-K] 에 맞춰본 결과 알려진 공식과 상당히 일치하는 것을 확인할 수 있었으며 아미노산의 조성도 시스테인 (Cys) 이 약 1/3 정도 함유하는 사실을 확인 할 수 있었다. BLAST 결과를 토대로 선정 되어진 80개의 참고 서열 중 아미노산 레벨에서 가장 높은 스코어로 align 되는 서열들은 담수패인 Dreissena polymorpha (zebra mussel), Unio tumidus (swollen river mussel) and Crassostrea ariakensis (suminoe oyster) 등으로 나타났다. ClustalX 를 통해 multiple align 한 후 Neighbor-Joining 방법에 따라 phylodendrogram을 그려본 결과 Pisidium (Neopisidium)coreanum의 MT는 Dreissena polymorpha (Dp), Crassostrea gigas (Cg4), Crassostrea virginica (Cv7) 등의 생물들과 같은 군으로 묶이는 것을 관찰 할 수 있었다. Psipred 소프트웨어를 통해 2D 구조를 비교 분석 한 결과도 multiple align 및 phylodendrogram 결과와 밀접한 관계가 있음을 알 수 있었다. 이러한 결과를 통해 EST를 통해 밝혀진 Pisidium (Neopisidium) coreanum의 MT 서열은 근연종 들의 서열과 일치함을 알 수 있었으며 이러한 결과에 기인하여 MT 서열은 분류에 사용 될 수 있다고 사료된다.

Keywords

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