市售14份蛇粗毒的16S rRNA基因序列分析

Analysis of 16S rRNA gene sequences from fourteen snake venoms bought from market

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16SrRNA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。

Objective：Extraction of genomic DNA from dried snake venoms of different resources and inferring their real original species by analysis of their partial 16S gene sequences. Methods：Extract the genomic DNA from fourteen snake venoms which labeled from 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 compared against 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 straight- forward method for verifying the identity of animal venoms.