The Korean Journal of Malacology (한국패류학회지)

The Malacological Society of Korea (한국패류학회)
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Identification and in silico analysis of two types of serpin genes from expressed sequence tags (ESTs) of the Oriental land snail, Nesiohelix samarangae

동양달팽이 (Nesiohelix samarangae) 의 expressed sequence tags (ESTs) 로부터 분리한 2종류의 Serpin 유전자 분석

  • Park, So Young (Department of Pathology, College of Medicine, Soonchunhyang University) ;
  • Jeong, Ji Eun (Department of Applied Biology College of Natural Resources and Life Science, Dong-A University) ;
  • Hwang, Hee Ju (Department of Life Science, Soonchunhyang University) ;
  • Wang, Tae Hun (Department of Life Science, Soonchunhyang University) ;
  • Park, Eun Bi (Department of Life Science, Soonchunhyang University) ;
  • Kim, Yong Min (Korea National Park Service, Taeanhaean National Park Office) ;
  • Lee, Jun-Sang (Institute of Environmental Research, Kangwon National University) ;
  • Han, Yeon Soo (Division of Plant Biotechnology, College of Agriculture and Life Science, Chonnam National University) ;
  • Yang, Seung-Ha (Department of Pathology, College of Medicine, Soonchunhyang University) ;
  • Lee, Yong Seok (Department of Life Science, Soonchunhyang University)
  • 박소영 (순천향대학교 의과대학 병리학교실) ;
  • 정지은 (동아대학교 생명자원과학대학 응용생물공학과) ;
  • 황희주 (순천향대학교 자연과학대학 생명시스템학과) ;
  • 왕태훈 (순천향대학교 자연과학대학 생명시스템학과) ;
  • 박은비 (순천향대학교 자연과학대학 생명시스템학과) ;
  • 김용민 (국립공원관리공단 태안해안국립공원사무소) ;
  • 이준상 (강원대학교 환경연구소) ;
  • 한연수 (전남대학교 농업생명과학대학 식물생명공학부) ;
  • 양승하 (순천향대학교 의과대학 병리학교실) ;
  • 이용석 (순천향대학교 자연과학대학 생명시스템학과)
  • Received : 2014.06.14
  • Accepted : 2014.06.27
  • Published : 2014.06.30
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Abstract

Serpins are a group of proteins involved in the regulation of serine and other type of proteases, and have been identified in many kinds of organisms from invertebrates to vertebrates. Serpins are known to regulate the proteolytic cascades of the innate immune pathways in addition to their roles in blood coagulation, angiogenesis, fibrinolysis, inflammation and tumor suppression. In this study, we have isolated two partial serpin gene fragments from expressed sequence tags (ESTs) of Nesiohelix samarangae. Dotplot analysis indicates that they are of two different types, Ns-serpin type 1 and Ns-serpin type 2. Ns-serpin type 1 has 819 bp coding region (272 amino acids), whereas Ns-serpin type 2 has 555 bp coding region (185 amino acids). Molecular phylogenetic analysis shows that the identified serpins have high similarities to their counterparts in the California see slug, Aplysia californica. Yet, the precise biological and immunological roles of these Ns-serpins remain to be further investigated using RNA interference and other molecular techniques.

동양달팽이의 EST 에서 serpin 유전자는 2가지 type이 추정되었다. Ns-serpin type 1 (partial) 서열의 코딩 영역은 총 819 bp 이었으며, 272개의 아미노산으로 이루어져 있었으며, Ns-serpin type 2 (partial) 의 코딩영역은 총 555 bp, 185개의 아미노산으로 이루어져 있었다. Laminarin 처리 전에는 Ns-serpin type 1이 2개 발현되었으며 처리 후에는 Ns-serpin type 2가 1개 발현되고 있었다. BLAST 결과를 바탕으로 유사도가 높은 17개의 참고 서열과 동양달팽이의 Ns-serpin type 1, 2의 아미노산 서열을 MEGA6 프로그램의 clustalW 엔진을 이용하여 multiple sequence alignment 를 수행한 결과, 척색동물의 포유강 9종, 조류강 1종, 양서강 1종과 척색동물인 해초강, 절지동물의 거미강 1종과 곤충강 1종이 같은 군으로 묶였으며, 연체동물문의 경우 동양달팽이의 두 가지 type 을 포함한 복족강 4종이 같은 군으로 묶이는 것을 확인하였다. Psipred 프로그램을 활용하여 예측한 2D 구조도 multiple align 및 phylodendrogram 결과와 연관이 있음을 확인할 수 있었다. EST를 통해 밝힌 동양달팽이의 두 가지 type의 Ns-serpin 서열은 Aplysia californica 등의 근연종들의 서열과 유사하였다. 본 연구에서는 무척추동물에서의 선천성 면역과 연관이 있는 것으로 잘 알려진 두 종류의 serpin 유전자 서열을 동양달팽이 EST 결과에서 발굴 하였으며, 동양달팽이에서도 여러 가지 type의 serpin 이 존재 할 수 있음을 입증하였다.

Keywords

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  1. Molecular Phylogenetic Study of Nesiohelix samarangae Based on CO-I Gene vol.30, pp.4, 2014, https://doi.org/10.9710/kjm.2014.30.4.391