市售14份蛇粗毒的16S_rRNA基因序列分析___

摘要   目的:从14份标签上注明的来自不同地区的4种干燥蛇粗毒——分别来自舟山眼镜蛇、孟加拉眼镜蛇、尖吻蝮蛇和短尾蝮蛇中提取总DNA，并结合PCR和序列分析技术对粗毒的来源进行核实。方法:分别从上述粗毒中提取总DNA，同时以4种对应蛇肌肉组织中提取的总DNA作为对照，用1对线粒体16S rRNA基因通用引物16H1/16L1进行扩增。结果:测序结果提交NCBI，并与GenBank中的同源序列进行比对和构建进化树。结论:序列分析结果证实扩增的均为正确的来自对应蛇种的线粒体16S基因序列，该技术有可能推广用于对各种动物毒液类毒素样品来源的身份进行核实。

关键词：蛇毒；物种鉴定；16S rRNA基因；DNA测序

Abstract   Objective:Extraction of genomic DNA from dried snakevenoms of different resources and inferring their real original species by analysis of their partial 16S gene sequences.Methods:Extractthe genomic DNAfrom fourteen snake venoms which labeledfrom different proviences in China and Thailand.These venoms are from Naja atra,Naja kaouthia,Deinagkistrodon acutus and Gloydius brevicaudus,respectively.DNA extracted from the muscles were used as controls.Their 16S gene sequences were amplified and sequenced.Results:All the sequences were submitted to NCBI and comparedagainst their homologous sequences in the GenBank database.The phylogenetic trees were constructed by Neighbor-joining(NJ) and Maximum parsimony(MP) method.Conclusion:The phylogenetic analysis concluded that they were from the corresponding snake species.This approach can offer an straightforward method for verifyingthe identity of animal venoms.

Key words:snake venom;species identification;16S rRNA gene;DNA sequencing