Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Application for Identification of Food Raw Materials by PCR using Universal Primer

일반 프라이머를 이용한 PCR의 식품원료 진위 판별에 적용

  • Park, Yong-Chjun (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Jin, Sang-Ook (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Lim, Ji-Young (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) ;
  • Han, Sang-Bae (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) ;
  • Lee, Kwang-Ho (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Yoon, Hae-Seong (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration)
  • 박용춘 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 진상욱 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 임지영 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 김규헌 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이재황 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 조태용 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이화정 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 한상배 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이상재 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이광호 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 윤혜성 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀)
  • Received : 2012.03.05
  • Accepted : 2012.09.17
  • Published : 2012.09.30
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Abstract

In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

본 연구는 식품원료의 진위여부를 판별하기 위한 시험법으로 일반 프라이머를 이용한 DNA barcode 기법을 도입하였다. 동물성식품원료의 경우 미토콘드리아 DNA 중 cytochrome oxidase subunit I(COI) 부위 검출을 위하여 디자인된 프라이머(LCO1490/HCO2198 및 VF2/FISH R2)와 cytochrome b(cyt b) 부위 검출을 위하여 디자인된 프라이머(L14724/H15915)를 사용하였다. 상기 3 종류의 프라이머를 사용하여 가축류 6종(소, 돼지, 염소, 양, 말 및 사슴), 가금류 4종(닭, 오리, 칠면조 및 타조), 어류 7종(명태, 대구, 청대구, 청어, 송어, 다랑어 및 우럭)을 대상으로 PCR 후 전기영동하여 예상되는 PCR 산물의 생성 유무를 확인하였다. 가축류 6종에 대하여는 LCO1490/HCO2198, VF2/FISH R2 및 L14724/H15915 프라이머를 사용한 경우 COI 및 cyt b가 모두 검출되었으며, 가금류 4종은 LCO1490/HCO2198 및 VF2/FISH R2 프라이머를 사용한 경우만 COI이 검출되었다. 또한 어류 7종은 VF2/FISH R2 프라이머를 사용한 경우에만 COI 부위가 검출됨을 확인하였다. 식물의 경우 엽록체 DNA 부위를 이용하여 디자인된 3 종류의 프라이머(trnH/psbA, rpoB 1F/4R 및 rbcL 1F/724R)를 사용하였다. 각각의 프라이머를 이용하여 식물 5종(마늘, 양파, 무, 녹차 및 시금치)에 대하여 실험한 결과 3종류의 프라이머에서 PCR의 산물을 모두 확인하였으며 trnH/psbA 프라이머의 경우 식물 종마다 PCR 산물의 크기는 다르게 검출되었다. 본 연구에서는 17종의 식품원료별 일반 프라이머 및 PCR 조건을 확립하였으며, 생산된 PCR 산물을 대상으로 염기서열을 결정하고 유전자은행에 있는 염기서열과 DB 비교 분석을 통하여 식품에 사용된 원료의 진위여부 판별에 적용이 가능할 것으로 기대된다.

Keywords

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