주요 법생물 대상의 DNA 바코드 분석 연구

Species identification of wild animals is critical in a wide range of forensic casework, including bush meat harvesting, unregulated trade in protected species or species derivatives, unauthorized introduction of exotic species, and food fraud. The most traditional method for species identification and taxonomy has been morphological trait analysis. When morphological identification fails, genetic identification based on the DNA barcoding methods can be used to match sequences from unknown samples to sequences from a reference data. Although DNA barcoding technology is currently being used to solve various forensic casework of wild animals, it is difficult to adopt as the scientific evidence because standardized experiments and analysis methods are not applied. Therefore, an analysis guideline based on DNA barcode information and the phylogenetic basis of wild animals was developed in this study, and a validation experiment was carried out. To perform DNA barcoding, an appropriate molecular marker must be chosen, and the mitochondrial COI gene is most commonly used as a DNA marker in the case of wild animals. Furthermore, it can be considered very useful as barcode information because the mitochondrial COI gene accumulates a much greater amount of genetic information than other genes. The mitochondrial COI sequences have been stacked in public databases with a vast amount of information, but due to biological complexity that may occur within the ecosystem, incorrect information is frequently provided. Therefore, prior to the utilization of DNA barcode information, a procedure for verifying errors must be preceded. In addition to gene sequence similarity analysis, the analysis step of reinterpreting the species as a molecular phylogenetic unit (MOTU) through phylogenetic analysis and checking the barcode gap is needed. In this study, step-by-step guidelines for DNA barcode analysis were prepared based on the biological characteristics of wild animals, and COI genetic data verification of major wild animals in the Korean Peninsula was performed using the DNA barcode analysis method. As a result, the intraspecific variation based on the COI gene of major wild animals on the Korean Peninsula was found to be within 1% on average and within 2% of the maximum, which was confirmed to be statistically significant: barnacles (avg. 0.9%), blowflies (avg. 0.35%), cartilaginous fishes (avg. 0.34%), falconiformes (avg. 0.32%), waterfowl (avg. 0.29%), passerines (avg. 0.35%), artiodactyls (avg. 0.54%), and carnivores (avg. 0.50%). In addition, the DNA barcoding method of wild animals was conducted to confirm the tester's proficiency and the effectiveness of the analysis method, and 100% reproducibility was secured in the intra/inter laboratory reproducibility and repeatability test.