线粒体16SrRNA和Cytb基因复合扩增进行种属鉴定

目的建立一种用于种属鉴定的线粒体DNA16SrRNA基因和细胞色素b基因荧光标记复合扩增检测体系。方法利用引物设计软件Primer5.0对mtDNA序列的16SrRNA基因和细胞色素b基因各设计一对引物,建立复合扩增体系,分别扩增人和牛、猪、狗、鸡、草鱼5种常见动物,用310遗传分析仪对产物进行分析。结果人和5种动物DNA扩增产物均出现两个峰,Cytb通用引物的扩增产物为人与动物的共有峰,为358bp;16SrRNA基因的扩增产物为人与动物间存在位置差异的特异峰,位于231～256bp之间。结论该复合扩增体系可以明确区分人和5种动物样本,可用于种属鉴定。

Multiple Amplification of 16SrRNA Gene and Cytb Gene in Mitochodrial DNA for Species Identification

Objective To establish a fluorescent multiple amplification system of 16SrRNA and Cytb genes located in mitochondrial DNA for species identification.Methods A pair of primers of 16SrRNA gene and Cytb gene of the mitochondrial DNA was designed with the software Primer 5.0 to construct a multiple amplification system.The amplified products from human and five species of animals,including cattle,pig,dog,chicken and grass carp were analyzed by 310 Genetic Analyzer.Results The amplified products of these samples showed two peaks.The common one was 358 bp and the specific one different in unique species was between 231 bp and 256 bp.Conclusion The multiplex amplification system can exactly distinguish the species of human from five common animals.