Andrejevic Marko, Markovic Milica Keckarevic, Bursac Biljana, Mihajlovic Milica, Tanasic Vanja, Kecmanovic Miljana, Keckarevic Dusan
DNA Centre for Genetics, Belgrade, 11000, Serbia.
Faculty of Biology, Center for Forensic and Applied Molecular Genetics, University of Belgrade, Studentski trg 3, Belgrade, 11000, Serbia.
Forensic Sci Med Pathol. 2019 Jun;15(2):169-177. doi: 10.1007/s12024-019-00096-4. Epub 2019 Feb 26.
Mitochondrial DNA (mtDNA), especially the gene for cytochrome b (MT-CYB), has been found to be highly informative for species identification. In this study, we present the results of the analysis of a 127 bp long fragment of MT-CYB, amplified using universal primers, variable enough to be used for species identification and discrimination, even in highly degraded animal samples. The total number of analyzed species in this study was 30, including 17 mammalian and 13 bird species. Using a newly created primer pair, we successfully amplified and sequenced the target sequence in almost all tested species. The amplification was incomplete in just two species, and as a result, partial, but still variable sequences, were obtained. Using the target fragment we successfully identified all tested samples. Initial results suggested that the intraspecies genetic diversity of the target region, in all tested species, was low - from 0 to 4.72%. The interspecies genetic diversity of the target region, crucial for successful discrimination, showed relatively high diversity, ranging from 8.36% to 42.52%. Given its short length, the target region should be used for species determination, particularly in samples that are degraded or are low in DNA quantity.
线粒体DNA(mtDNA),尤其是细胞色素b基因(MT - CYB),已被发现对物种鉴定具有高度信息价值。在本研究中,我们展示了使用通用引物扩增得到的MT - CYB基因127bp长片段的分析结果,该片段具有足够的变异性,即使在高度降解的动物样本中也可用于物种鉴定和区分。本研究中分析的物种总数为30种,包括17种哺乳动物和13种鸟类。使用新创建的引物对,我们成功地在几乎所有测试物种中扩增并测序了目标序列。仅在两个物种中扩增不完全,因此获得了部分但仍具有变异性的序列。使用目标片段,我们成功鉴定了所有测试样本。初步结果表明,在所有测试物种中,目标区域的种内遗传多样性较低,为0至4.72%。对于成功区分至关重要的目标区域的种间遗传多样性显示出相对较高的多样性,范围为8.36%至42.52%。鉴于其长度较短,目标区域应用于物种鉴定,特别是在降解或DNA含量低的样本中。