한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제43권3호
  • /
  • Pages.296-299
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  • 2015
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  • 1598-642X(pISSN)
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  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
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Cowpea mild mottle virus 특이유전자 검출을 위한 검역진단시스템 개발

Development of a Diagnostic System for the Detection of the Cowpea mild mottle virus Specific Gene in Quarantine

  • 이시원 (국립환경과학원 상하수도연구과) ;
  • 이진영 (단국대학교 자연과학대학 미생물학과) ;
  • 문보영 (농림축산검역본부 호남지역본부 시험분석과) ;
  • 김창수 (국립환경과학원 상하수도연구과) ;
  • 신용길 (농림축산검역본부 인천공항지역본부 시험분석과) ;
  • 노재영 (단국대학교 자연과학대학 미생물학과)
  • Lee, Siwon (Water Supply & Sewerage Research Division, National Institute of Environmental Research) ;
  • Lee, Jin-Young (Department of Microbiology, College of Natural Sciences, Dankook University) ;
  • Moon, Bo Yeong (Experiment & Analysis Division, Honam Regional Office, Animal and Plant Quarantine Agency) ;
  • Kim, Chang Soo (Water Supply & Sewerage Research Division, National Institute of Environmental Research) ;
  • Shin, Yong-Gil (Experiment & Analysis Division, Incheon International Airport Regional Office, Animal and Plant Quarantine Agency) ;
  • Rho, Jae-Young (Department of Microbiology, College of Natural Sciences, Dankook University)
  • 투고 : 2015.07.20
  • 심사 : 2015.08.29
  • 발행 : 2015.09.28
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초록

Cowpea mild mottle virus (CPMMV)는 국내 미보고 바이러스로, 넓은 숙주범위를 가지며, 국내 관리급 검역바이러스로 지정되어 있다. 본 연구에서는 검역현장에서 신속하게 CPMMV를 진단할 수 있는 방법을 개발하였다. 이번 연구는 사용자를 위한 검사방법 개발을 위하여, 기존의 검역현장에서 활용하는 PCR 조성과 조건을 활용하였다. 본 연구에서는 CPMMV를 특이적으로 진단할 수 있는 2개의 RT-PCR 프라이머를 개발하였으며 각각 579와 638 bp를 증폭할 수 있다. 최종적으로 증폭되는 nested PCR 산물의 크기는 (579 → 298 bp)과 (638 → 252 bp)로 CPMMV의 특이 유전자를 진단할 수 있다. 또한, 제한효소 Xho I이 반응할 수 있는 염기 서열을 삽입하여 양성대조구 플라스미드를 개발하였다. 이것은 실험실 오염으로부터 거짓양성을 검증할 수 있게 하였다. 본 연구는 CPMMV와 관련된 수입 작물로부터 검역진단에 기여할 것이라고 사료된다.

Cowpea mild mottle virus (CPMMV) has a wide range of hosts, such as the pea family and tomato. CPMMV is a non-reported virus in Korea, and is domestically designated as a controlled virus associated with plant quarantine. In this study, a rapid diagnostic method for the detection of CPMMV at quarantine sites was developed. For the development of a user-based system, the PCR compositions and conditions use existing methods of quarantine for the viruses. Two sets of RT-PCR and nested PCR were developed in this study that could be amplified from 579 → 298 dp and 638 → 252 bp, respectively. Furthermore, a sequence inserted positive control plasmid was developed, which is able to identify false-positives resulting from laboratory contamination. The findings of this study are important for the diagnosis of CPMMV in imported crops held in plant quarantine.

키워드

참고문헌

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