Journal of the Korea Academia-Industrial cooperation Society (한국산학기술학회논문지)

The Korea Academia-Industrial cooperation Society (한국산학기술학회)
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Expression of the red sea bream iridovirus (RSIV) capsid protein using a yeast surface display method

효모표면표출(YSD) 기법을 이용한 참돔 이리도바이러스(RSIV) 외피단백질의 발현

  • Suh, Sung-Suk (South Sea Environment Research Department, Korea Institute of Ocean Science & Technology) ;
  • Park, Mirye (South Sea Environment Research Department, Korea Institute of Ocean Science & Technology) ;
  • Hwang, Jinik (South Sea Environment Research Department, Korea Institute of Ocean Science & Technology) ;
  • Lee, Taek-Kyun (South Sea Environment Research Department, Korea Institute of Ocean Science & Technology)
  • 서승석 (한국해양과학기술원 남해특성연구부) ;
  • 박미례 (한국해양과학기술원 남해특성연구부) ;
  • 황진익 (한국해양과학기술원 남해특성연구부) ;
  • 이택견 (한국해양과학기술원 남해특성연구부)
  • Received : 2014.06.23
  • Accepted : 2014.08.07
  • Published : 2014.08.31
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Abstract

The red seabream iridovirus (RSIV), which belongs to the iridoviridae, causes infectious fish diseases in many Asian countries, leading to considerable economic losses to the aquaculture industry. Using the yeast surface display (YSD) technique, a new experimental system was recently developed for the detection and identification of a variety of marine viruses. In this study, a coat protein gene of RSIV was synthesized based on the nucleotide sequence database and subcloned into the yeast expression vector, pCTCON2. The expression of viral coat proteins in the yeast strain, EBY100, was detected by flow cytometry and Western blot analysis. Finally, they were isolated from the yeast surface through a treatment with ${\beta}$-mercaptoethanol. The data suggests that the YSD system can be a useful method for acquiring coating proteins of marine viruses.

참돔 이리도바이러스(RSIV)는 이리도바이러스과에 속하며, 많은 아시아 국가에서 감염성 어류 질병을 유발하여 양식산업에 커다란 경제적 손실을 입히는 바이러스이다. 우리는 최근에 효모표면발현(yeast surface display, YSD)를 사용하여 다양한 해양바이러스를 동정하고 검출할 수 있는 새로운 실험시스템을 개발하였다. 이 연구에서 우리는 참돔 이리도 바이러스(RSIV)의 외피단백질을 효모표면 발현 기법을 이용하여 발현시켰다. 바이러스 외피단백질 유전자는 염기서열 데이터베이스에 기초하여 합성되었고, 효모발현벡터인 pCTCON2으로 서브클로닝되었다. 이 벡터는 효모 strain EBY100으로 형질전환 되었다. Flow cytometry와 Western blot analysis를 통해 RSIV 외피단백질의 발현을 확인하였다. ${\beta}$-mercaptoethanol 처리에 의해 발현된 바이러스 외피단백질을 효모 표면로부터 분리하였다. 이 연구의 결과는 YSD 시스템이 해양바이러스 외피단백질을 획득하기 위한 매우 좋은 발현시스템이라는 것을 보여준다.

Keywords

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