Differentiation of Cannabis subspecies by THCA synthase gene analysis using RFLP |
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| Affiliation: | 1. Institute for Medical Research, University of Belgrade, Belgrade, Serbia;2. Center for Forensic and Applied Molecular Genetics, Faculty of Biology, University of Belgrade, Belgrade, Serbia;3. Institute of Forensic Medicine, Clinical Center of Vojvodina, Faculty of Medicine, University of Novi Sad, Novi Sad, Serbia;4. Institute for forensic medicine, criminology and medical deontology, Medical Faculty, University of St. Cyril and Methodius, Skopje, Macedonia;5. Faculty of Mathematics, University of Belgrade, Belgrade, Serbia;6. Institute of History, Belgrade, Serbia;1. Centro Regionale Antidoping e di Tossicologia “A. Bertinaria”, Orbassano (TO), Italy;2. Forensic Genetics Laboratory - Legal Medicine Unit Department of Medicine, University of Barcelona, Spain;3. Department of Biology, University of Florence, Italy;4. Reparto CC Investigazioni Scientifiche di Roma, Sezione di Biologia, Roma, Italy;5. Forensic Genetics Laboratory, Ceinge-Federico II University of Naples, Italy;6. Department of Chemistry, University of Torino, Italy |
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| Abstract: | Cannabis sativa subspecies, known as industrial hemp (C. sativa sativa) and marijuana (C. sativa indica) show no evident morphological distinctions, but they contain different levels of psychoactive Δ-9-tetrahidrocanabinol (THC), with considerably higher concentration in marijuana than in hemp. C. sativa subspecies differ in sequence of tetrahydrocannabinolic acid (THCA) synthase gene, responsible for THC production, and only one active copy of the gene, distinctive for marijuana, is capable of producing THC in concentration more then 0,3% in dried plants, usually punishable by the law.Twenty different samples of marijuana that contain THC in concentration more then 0,3% and three varieties of industrial hemp were analyzed for presence of an active copy of THCA synthase gene using in-house developed restriction fragment length polymorphism (RFLP) method All twenty samples of marijuana were positive for the active copy of THCA synthase gene, 16 of them heterozygous. All three varieties of industrial hemp were homozygous for inactive copy.An algorithm for the fast and accurate forensic analysis of samples suspected to be marijuana was constructed, answering the question if an analyzed sample is capable of producing THC in concentrations higher than 0.3%. |
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| Keywords: | Cannabis Hemp False-negative THC RFLP Algorithm |
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