利用线粒体DNA Cyt b基因PCR-RFLP分析方法鉴别羊肉和鸭肉

Identification of mutton from duck meat by PCR-RFLP analysis of mitochondrial DNA Cyt b gene

建立了一种利用线粒体DNA(mtDNA) Cyt b基因PCR-RFLP分析来鉴别羊肉和鸭肉的方法。采用一对通用引物扩增绵羊、山羊和鸭的mtDNACytb基因,并对扩增产物用DNA限制性内切酶Bsu36I和SpeI进行酶切,电泳分析酶切产物的变化。结果表明通用引物可扩增羊和鸭472bp的PCR产物,经两种内切酶酶切后,绵羊、山羊和鸭的PCR产物分别被切为大小不同的片段,其中绵羊和山羊被SpeI切为213bp和259bp,而鸭则被Bsu36I切为95bp和377bp。利用PCR-RFLP分析mtDNA Cyt b基因的方法操作简单,是一种快速鉴别羊肉和鸭肉的可靠方法。

A method to distinguish mutton of sheep and goat from duck meat was developed based on the PCR-RFLP analysis of mitochondrial DNA（mtDNA） Cyt b gene. A pair of universal PCR primers was employed to amplify a fragment of mtDNA Cyt b gene in sheep,goat and duck. The PCR products were further digested by two DNA restriction endonucleases of Bsu361 and Spel. The results showed that the primers amplified successfully a 472bp long DNA fragment from all three species. The PCR-RFLP patterns resulted from Spel digestion of both sheep and goat PCR products carried two bands in 213bp and 259bp which were different from that of duck PCR product being digested by Bsu361 into 95bp and 377bp. This study demonstrated that the PCR-RFLP analysis of mtDNA Cyt b gene was a rapid method to accurately distinguish mutton of sheep and goat from duck meat.