关键词: 物种鉴定, 细胞色素b 基因, 分子遗传学, 小麂

Abstract:

For a long time, illegal poaching was a big challenge in wildlife conservation. In some cases, in which unambiguous morphological evidence was absent, it was difficult to identify the correct species, so the judicial process was impeded. In the present study, identification of species is possible by DNA sequencing of the material, especially a partial DNA sequence of the cytochrome b gene. Four cooked muscle samples that were suspected as coming from wildlife were collected from four different cities. DNA was extracted from the muscle samples, then a fragment of cytochrome b was amplified with the universal primers L14724 and H15149. PCR products were purified and sequenced,and four sequences of 393 to 405 bp were determined. They shared the same haplotype. After blasting, sequences were compared with the sequences of eight related species registered in GenBank, and the phylogenetic tree was constructed by the neighbor-joining method. The results of genetic distance (K2P) analysis showed that the genetic distance between the unknown species and different species ranged from 0.00 (unknown samples vs Muntiacus reesesi)to 0.106 (vs Cervus nippon). At the same time, these unknown samples of animal remains were identified by phylogenetic analysis with neighbor-joining tree, which shows that the samples and Muntiacus reeversi were clustered together in the system. On the basis of the case, some discussion was carried out on the key steps in the molecular genetic approach such as the choice of molecular markers and operational technique for molecular genetic diagnostic approach, and additionally, some conservation suggestions are put forward.

Key words: Cytochrome b gene, Molecular genetics, Muntiacus reeversi, Species identification