12S rRNA和Cyt b基因序列测定在獐乳制品鉴定中的应用

Application of Sequencing 12S rRNA and Cyt b Gene to Identification of Dairy Products of Hydropotes inermis

采用改良的蛋白酶K消化和酚/氯仿抽提的方法从脂肪含量很高的动物乳制品及胃组织中提取出基因组总DNA,利用特异引物扩增了线粒体12S rRNA基因和Cyt b基因的部分片段,测定了12S rRNA和Cyt b基因的PCR扩增产物序列,使用BLAST搜索软件将其序列与GenBank中的同源序列进行比对,并利用DNAMAN分析软件分析序列同源性。结果表明3件检材均来源于獐Hydropotes inermis。本研究所用方法在野生动物乳制品鉴定中具有较高的应用价值。

A modified method of DNA extraction was applied to extract DNA fi＇om animal dairy products and stomach tissue, which containing high fat. Polymerase chain reaction （PCR） assays were performed using specific primers to a part of mitoehondrial 12S rRNA gene and Cyt b gene. The PCR products of mitoehondrial 12S rRNA gene and Cyt b gene were sequenced. And the obtained sequences were aligned with homologous sequences derived from GenBank by BLAST and DNA- MAN software. The results indicated that all of the samples were derived from Hydropotes inermis. This study provids a good example for the identification of animal daily products.