线粒体DNA cytb和HVI的分析用于种属鉴定

目的以线粒体DNA为目标序列,探讨生物检材的种属来源问题。方法复合扩增线粒体DNA细胞色素b基因(Cb)片段和D-环HVI上人源特异性DNA片段,2%琼脂糖凝胶电泳检测复合扩增产物谱带;用常规测序技术获得种属来源不明的检材Cytb基因序列,登陆美国国家生物信息中心网站主页(http://www.ncbi.nlm.nih.gov),将Cytb序列的测序结果用BLAST2.2.92004.5.1]进行匹配查询,查询数据库中存在的与其相匹配的物种条目。结果检材经复合扩增后电泳检测可区分人源性检材和非人源性检材;用生物信息法可确定检材种属来源结论检测线粒体DNA细胞色素b基因和D-环HVI的有关序列可在DNA分子水平上鉴别人源性检材和非人源性检材,结合测序的分子生物信息学方法,可对检材进行种属鉴定。

Analysis of cytb and HVI in Mitochondrial DNA for Species identification

Objective To study species identification of forensic biological specimens from the MtDNAsequence. Methods Fifteen species(human, cow, sheep, pig, fish, chicken, duck, dog,chimpanzee, monkey, panda, deer, lion, leopard and zebra) were concerned. Duplex amplification of cytb and HVI were carried out and the PCR products were analyzed by agarose gel electrophoresis. The cytb gene was sequenced and inquired. results Human and nonhuman samples could be simply distinguished by electrophoresis bands of cytb and HIVI PCR products, the species identification could be finished by inquiring cytb sequence matching on Internet(http://www.ncbi.nlm.nih.gov). Conclusion From the results obtained, it was shown that the mtDNAtesting system could be a valuable tool for species identification.