Peppin Lindsay, McEwing Ross, Carvalho Gary R, Ogden Rob
Molecular Ecology and Fisheries Genetics Laboratory, School of Biological Sciences, 3rd Floor, Environment Centre Wales, Bangor University, Bangor, Gwynedd LL57 2UW, UK.
J Forensic Sci. 2008 Nov;53(6):1358-62. doi: 10.1111/j.1556-4029.2008.00857.x. Epub 2008 Aug 25.
Attempts to prevent illegal trade in bile and gallbladders from Asiatic black bears, Ursus thibetanus, are hampered by difficulties associated with identifying such items. We extracted DNA from bile crystals of unknown species origin and generated partial cytochrome b (cyt b) sequences using either universal primers (positioned in conserved regions of cyt b), or primers designed on existing U. thibetanus sequences (UT). Species origin was determined by aligning resolved sequences to reference sequence data. The universal primers were unsuitable for U. thibetanus identification when multiple species templates were present in the samples. The UT primers amplified U. thibetanus DNA from all sample extracts, including those containing mixed species templates. The amplified fragment can distinguish U. thibetanus from the most closely related species, U. americanus, a distinct advantage of DNA sequencing over the methods currently used to analyze suspected U. thibetanus bile.
防止亚洲黑熊(Ursus thibetanus)胆汁和胆囊非法贸易的努力因难以识别此类物品而受阻。我们从未知物种来源的胆汁晶体中提取DNA,并使用通用引物(位于细胞色素b(cyt b)的保守区域)或根据现有西藏棕熊序列(UT)设计的引物生成部分细胞色素b序列。通过将解析的序列与参考序列数据进行比对来确定物种来源。当样品中存在多个物种模板时,通用引物不适用于西藏棕熊的鉴定。UT引物能从所有样品提取物中扩增出西藏棕熊的DNA,包括那些含有混合物种模板的提取物。扩增片段可以将西藏棕熊与亲缘关系最近的物种美洲黑熊区分开来,这是DNA测序相对于目前用于分析疑似西藏棕熊胆汁的方法的一个明显优势。