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Development of Detection Method for Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus using 16S rRNA Gene

16S rRNA를 이용한 다금바리, 자바리, 능성어 판별법 개발

  • Park, Yong-Chjun (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Jung, Yong-Hyun (Food Standardization Division, Food & Drug Administration) ;
  • Kim, Mi-Ra (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Shin, Joon-Ho (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Kim, Kyu-Heon (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Lee, Jae-Hwang (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Cho, Tae-Yong (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Lee, Hwa-Jung (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Lee, Sang-Jae (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Han, Sang-Bae (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration)
  • 박용춘 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 정용현 (식품의약품안전청 식품기준과) ;
  • 김미라 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 신준호 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 김규헌 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이재황 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 조태용 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이화정 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이상재 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 한상배 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀)
  • Received : 2012.07.03
  • Accepted : 2012.12.17
  • Published : 2013.02.28

Abstract

Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus are involved in the Perciformes Order and Serranidae Family. When E. bruneus and E. septemfasciatus are fully grown, the striped pattern on the body gradually disappears. Therefore, morphological classification of adult fishes is quite difficult to identify the differences to N. spinosus. In this study, we investigate the method to differentiate those using PCR. To design the primers, 16S rRNA region of N. spinosus, E. bruneus, and E. septemfasciatus registered in the GeneBank (www.ncbi.nlm.nih.gov) have been used and for the analysis, Bio Edit ver. 7.0.9.0 was used. As a result, it was design NS-003-F/NS-005-R (136 bp), EB-001-F/EB-002-R (181 bp), and ES-001-F/ES-001-R (123 bp) primers for the differentiation of each 3 different fishes. Therefore, the species-specific primer sets would be a useful tool for scientific and speedy differentiation against the illegal distribution for consumer protection.

다금바리, 자바리 및 능성어는 농어목(Perciformes Order) 바리과(Serranidae Family)에 속하며, 고급횟감으로 인기가 높은 어종이다. 자바리 및 능성어의 경우 몸통부분에 줄무늬가 선명하게 있으나 성어가 되면서 점차 불분명해지는 특징이 있어 무늬의 유무로 인하여 다금바리와 구별하기는 쉽지 않다. 또한 자바리는 제주도에서 다금바리라는 명칭으로 불리고 있으며, 일부 유통업자들이 자바리 및 능성어를 다금바리로 유통시키는 사례가 있어 종 특이 프라이머를 이용한 판별법을 마련하게 되었다. 종 특이 프라이머를 설계하기 위하여 유전자은행(www.ncbi.nlm.nih.gov)에 등록되어있는 다금바리(Accession No. AY947575), 자바리(Accession No. JN603832), 능성어(Accession No. AY947559), 우럭(Accession No. DQ678295) 등의 16S DNA부위의 염기서열을 대상으로 하였으며 비교 및 분석에는 소프트웨어인 BioEdit ver. 7.0.9.0 프로그램을 사용하였다. 그 결과 다금바리, 자바리, 능성어를 판별할 수 있는 각각의 NS-003-F/NS-005-R(136 bp), EB-001-F/EB-002-R(181 bp), ES-001-F/ES-001-R(123 bp) 프라이머를 개발하였으며, PCR 조건을 확립하였다. 또한 적용성 검토를 위하여 우럭, 참돔을 대상으로 실시한 결과 비 특이적 밴드가 형성되지 않는 것을 확인하였다. 따라서 본 연구에서 개발된 다금바리 등에 대한 판별법은 시중에 불법적으로 유통 가능성이 있는 제품을 신속하고 과학적으로 판별할 수 있어 식품안전관리에 활용도가 매우 클 것으로 기대된다.

Keywords

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