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目的应用mtDNA 16S rRNA基因测序方法进行肉产品种属鉴定。方法 15份有关部门送检的未知动物肌肉样本,3份市购猪、牛和羊的肌肉为对照样本。常规酚-氯仿法提取模板DNA,分别用通用引物和特异性引物扩增mtDNA的16S rRNA基因片段,产物用1.5%琼脂糖电泳检测,扩增的基因片段由上海生物工程公司进行序列测定及基因库同源性匹配查询。结果经通用引物扩增15份未知样本,均检测出阳性条带;用羊特异性引物扩增,无未知样本检出阳性条带;用猪及牛特异性引物扩增,分别有14份和1份样本检出阳性条带,经测序及同源性查询,分别与猪和牛的同源性为96%～100%。结论送检未知肉产品样本中14份是猪肉,1份是牛肉。
Objective To identify the species of meat products by using mtDNA 16S rRNA gene sequencing method. Methods Fifteen muscle samples of unknown animals submitted by relevant departments, three commercially available pigs, cattle and sheep muscles were used as control samples. The template DNA was extracted by conventional phenol-chloroform method. The mtDNA 16S rRNA gene fragment was amplified by universal primers and specific primers respectively. The product was detected by 1.5% agarose gel electrophoresis. The amplified gene fragment was sequenced by Shanghai Bioengineering Co., Library homology matches query. Results Fifteen unknown samples were amplified by universal primers, and the positive bands were detected. The positive bands were detected by using sheep-specific primers and no unknown samples were detected. The amplification products were 14 and 14 One sample showed a positive band, which was sequenced and homologous inquired. The homology was 96% ~ 100% with that of pig and cattle respectively. Conclusion 14 samples of unknown meat product samples were pork and 1 beef.