东南亚燕窝的12S rRNA和Cytb基因序列分析

Sequence Analysis of 12S rRNA and Cytb Genes from Bird's Nest in South-east Asia

关键词:  燕窝  DNA鉴定  12S核糖体核糖核酸  细胞色素b基因  DNA测序

Key words:  bird's nest  DNA identification  12S rRNA  Cytb  DNA sequencing

摘要:

目的 对实地采集的25份未进行任何加工的东南亚燕窝样品的12S rRNA和Cytb基因序列进行分析，确定东南亚地区燕窝来源物种的真实身份，其中包括2份珍贵的采集自马来西亚燕洞的黄燕和血燕标本。方法 25份未经过任何加工的样品采集自马来西亚、新加坡和越南地区的燕洞和燕屋，以直接提取自燕窝的基因组DNA为模板，利用重新设计的引物扩增得到12S rRNA和Cytb基因序列，并进行克隆测序。结果 测序结果与GenBank中的同源序列进行BLAST比对分析并构建亲缘关系树，结合样品采集地的金丝燕种类信息初步确定爪哇金丝燕（Aerodramus fuciphagus）和大金丝燕（Aerodramus maximus）。结论 分析结果证实扩增的均为正确的来自对应金丝燕属的线粒体基因序列，结合金丝燕种源地的物种分布资料和基因序列信息，为制定进口燕窝的质量标准提供了依据。

Abstract:

OBJECTIVE To confirm the real identity of imported bird's nest origin species in South-east Asia, the 12S rRNA and Cytb gene sequences from twenty-five bird's nest samples from South-east Asia were determined and analyzed, which including two precious bloody nest sample(yellow nest and bloody nest) collected from the birdnest cave in Malysia. METHODS Twenty five samples without any processing were collected from birdnest cave and birdnest house in Malaysia, Singapore and Vietnam. The genomic DNA directly extracted from bird's nest was used as template to amplify 12S rRNA and Cytb gene sequences by redesigned primers, and then cloned and sequenced. RESULTS All the sequences were BLAST against their homologous sequences in GenBank database and the gene trees were constructed. Based on the species information of swiftlets collected in the sample collection area, these samples were initially identified as Aerodramus fuciphagus and Aerodramus maximus. CONCLUSION The sequence aliment analyses approve that they are the right mitochondrial gene from the Aerodramus species. Combined with the species distribution data and gene sequence information of the original swiftlets, these sequence data acquired by this method can be used to formulate the quality standard of imported bird's nest.