The Korean Journal of Malacology (한국패류학회지)

The Malacological Society of Korea (한국패류학회)
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Identification, sequence characterization and expression analysis of the arginine kinase gene in response to laminarin challenge from the Oriental land snail, Nesiohelix samarangae

동양달팽이(Nesiohelix samarangae)의 arginine kinase 유전자 분석 및 발현 패턴에 관한 연구

  • Jeong, Ji Eun (Department of Life Science, Soonchunhyang University) ;
  • Lee, Yong Seok (Department of Life Science, Soonchunhyang University)
  • 정지은 (순천향대학교 자연과학대학 생명시스템학과) ;
  • 이용석 (순천향대학교 자연과학대학 생명시스템학과)
  • Received : 2013.08.29
  • Accepted : 2013.09.28
  • Published : 2013.09.30
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Abstract

Arginine kinase (ArK) is known to play an important role in most invertebrates the level of ATP by phosphorylation of phosphagens in cell and immuninty in living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.

동양달팽이의 arginine kinase 유전자는 염기서열 1065개로 이루어져있으며 355개의 아미노산으로 이루어져 있으며, BLAST 결과를 토대로 유사도가 높은 25개의 참고 서열과 동양달팽이의 arginine kinase의 아미노산 서열을 MEGA5 프로그램의 clustalW 모듈을 이용하여 multiple sequence alignment 를 수행한 결과, 연체동물문에 속하는 복족강 (5종), 두족강 (5종), 이매패강 (4종) 등에 속하는 생물들이 같은 군으로 묶였으며, 절지동물문 곤충강 에 속하는 나비목 (2종), 벌목 (1종), 노린재목 (2종) 등에 속하는 생물들이 같은 군으로 묶이고, 갑각강 (5종), 거미강 (1종) 에 속하는 생물들이 묶이는 것을 알 수 있었다. Psipred 소프트웨어를 통해 2D 구조를 비교 분석한 결과도 multiple align 및 phylodendrogram 결과와 밀접한 관계가 있음을 알 수 있었다. 시간에 따른 arginine kinase의 발현양상을 확인한 결과 control에 비하여 6시간에서 약 1.2배 정도 발현이 증가하는 것을 확인할 수 있었으며, 12시간이 지나면 점차 감소하는 것을 확인할 수 있었다. EST를 통해 밝혀진 N. samarangae 의 Ark 서열은 근연종들의 서열과 일치함을 알 수 있었으며, 본 연구 결과를 통해 무척추동물에서의 선천성 면역 관련 유전자 연구에 동양달팽이가 좋은 모델이 될 수 있음을 제시하고 있다.

Keywords

References

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