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Detection Method for Identification of Pueraria mirifica (Thai kudzu) in Processed Foods

가공식품 중 태국칡(Pueraria mirifica) 혼입 판별법 개발

  • Park, Yong-Chjun (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Jin, Sang-Wook (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Kim, Mi-Ra (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) ;
  • 진상욱 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 김미라 (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 : 2012.05.03
  • Accepted : 2012.12.03
  • Published : 2012.12.31

Abstract

In this study, ribulose bisphosphate carboxylase (rbcL), RNApolymeraseC (rpoC1), intergenic spacer (psbA-trnH), and second internal transcribed spacer (ITS2) as identification markers for discrimination of P. mirifica in foods were selected. To be primer design, we obtained 719 bp, 520 bp, 348 bp, and 507 bp amplicon using universal primers from selected regions of P. mirifica. The regions of rbcL, rpoC1, and psbA-trnH were not proper for design primers because of high homology about P. mirifica, P. lobata, and B. superba. But, we had designed 4 pairs of oligonucleotide primers from ITS2 gene. Predicted amplicon from P. mirifica were obtained 137 bp and 216 bp using finally designed primers SFI12-miri-6F/SFI12-miri-7R and SFI12-miri-6F/SFI12-miri-8R, respectively. The species-specific primers distinguished P. mirifica from related species were able to apply food materials and processed foods. The developed PCR method would be applicable to food safety management for illegally distributed products in markets and internet shopping malls.

식품 중 P. mirifica 원료 함유여부에 대한 판별법 마련을 위하여 식물의 종 동정에 일반적으로 사용되는 ribulose bisphosphate carboxylase (rbcL), RNA polymerase C (rpoC1), intergenic spacer (psbA-trnH) 및 second internal transcribed spacer (ITS2) 유전자부위를 선정하였다. 선정된 유전자부위를 증폭하기 위하여 일반 프라이머를 이용하였으며 각각 719 bp, 520 bp, 348 bp 및 507 bp의 PCR 산물을 확인하였다. 그리고 염기서열을 결정하고 유전자은행에 등록되어있는 염기서열과 유사성에 대한 분석한 결과 rbcL, rpoC1 및 psbA-trnH 부위는 상동성이 매우 높아 프라이머를 설계하기는 어려웠다. 그러나 ITS2의 경우 염기서열의 차이점이 있어 4종류의 프라이머를 설계하였다. 설계된 프라이머를 이용하여 P. mirifica, P. lobata, B. superba에 대한 PCR을 실시한 결과 SFI12-miri-6F/SFI12-miri-7R 및 SFI12-miri-6F/SFI12-miri-8R의 경우 P. lobata 와 B. superba에서 비 특이적 밴드가 없으며 P. mirifica에서 예상크기인 137 bp 및 216 bp를 확인할 수 있었다. 따라서 본 연구에서 개발된 P. mirifica을 판별할 수 있는 종 특이 프라이머는 식품가능원료 및 가공식품에 대한 적용할 수 있어 인터넷 쇼핑몰 등 시중에 불법적으로 유통되는 제품에 대한 안전관리에 활용도가 매우 클 것으로 기대된다.

Keywords

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