엽록체 DNA를 이용한 마약성 양귀비 판별법 개발
Development of Narcotic Poppy Identification Method using Chloroplast DNA
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초록
연구에서는 마약성 양귀비의 정확한 DNA 판별법 개발을 위해 양귀비속(genus Papaver) 식물 3 종(양귀비(P. somniferum L.), P. orientale L., 개양귀비(P. roheas L.))의 엽록체 유전체 염기서열을 Illumina Hiseq 2000을 이용하여 분석하고 마약성과 비마약성 사이의 특이적인 염기서열 차이를 발견하여 이를 바탕으로 PCR 산물의 길이 차이만으로 마약성 양귀비류와 비마약성 양귀비류를 간편하고 정확하게 구분할수 있는 마약성 양귀비 판별용 InDel (Insertions and deletions) 마커를 선정하였다. 3개 InDel 마커를 활용한 마약성 양귀비 판별법을 실제 감정물에 적용한 결과, 마약성분 테바인, 코데인, 그리고 몰핀이 검출된 샘플을 정확하게 식별하였다. 또한, 마약성분을 검출하기 어려운 미량의 씨앗, 외형이 훼손되거나 부패된 샘플, 그리고 여러 종류의 식물이 혼합되어 있는 샘플에서 마약성 양귀비를 판별할 수 있음이 확인되었다. 따라서 새롭게 선정된 InDel 마커를 이용한 마약성 양귀비 판별법은 GC-MS를 이용하는 마약 성분 검사법을보완하여 마약수사 실무에서 활용성이 높을 것으로 판단된다
Abstract
Among a variety of molecular markers, insertions and deletions (InDel) markers are proposed for the identification of narcotic opium poppies in this study. For the design of InDels markers, the chloroplast genome sequences of 3 papaver species (P. sominiferum, P. orientale, and P. rhoeas) were completed using Illumina Hiseq 2000 and the specific genetic differences among species were found. On the basis of the InDel size differences, three InDel markers were developed with the advantages of easy to set up and high accuracy to distinguish narcotic and non-narcotic poppies. As a result of applying the 3 InDel markers to the drug case samples, these markers correctly differentiated the samples contained the Thebaine, Codeine, and/or Morphine. In addition, it was confirmed that narcotic poppy could be identified in seeds without narcotic substances, in damaged or decomposed samples, and in samples containing a mixture of various plants. Therefore, the newly developed 3 InDel markers for the identification of narcotic poppies would be highly useful in drug investigations by complementing gas chromatography-mass spectrometry (GC-MS
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