COI序列应用于羊肉掺伪非定向筛查技术的研究

The Use of COI Sequence for Non-directional Screening to Detect Adulteration in Mutton

摘要:

本项目研究了应用COI序列对羊肉中掺入的其他动物源材料进行非定向筛查，并对山羊肉及绵羊肉进行分辨。利用了DNA条形码技术对六个羊品种（系）的线粒体基因组细胞色素酶基因片段进行引物筛选，并利用筛选出的引物对七种动物源样本的COI片段进行扩增，以确定所筛选出的引物是否能将羊肉与其他动物源样品进行有效区分。以LCOI490/HCO2198、F1-1/R1-1、ITS3/ITS4为引物，通过对36份血液样本（山羊3个品种，绵羊3个品种）的基因组DNA进行PCR扩增，其中以F1-1/R1-1为引物得到的PCR产物电泳检测条带清晰，结果重现性好；利用F1-1/R1-1引物对鸡肉、猪肉、驴肉、牛肉、莱芜黑山羊肉、洼地绵羊肉和鸭肉等七种样本的COI片段进行扩增，并对扩增片段进行测序，山羊、绵羊之间的序列同源性仅为36.62%，其他各品种间的序列差异也很明显。结果表明：以F1-1/R1-1为专用引物可以有效区分羊肉与其他动物源样品，并能够对山羊肉及绵羊肉样品进行有效区别。

Abstract:

This study investigated non-directional screening of other animal source materials incorporated in mutton by using COI sequence, while distinguishing goat meat and sheep meat. DNA barcoding technology was used to screen the mitochondrial genome cytochrome gene fragment of six sheep breeds (lines), and the selected primers were used to amplify the COI fragments of seven animal source samples to determine whether or not the primer can effectively distinguish the lamb from other animal source samples. Using LCOI490/HCO2198, F1-1/R1-1, and ITS3/ITS4 as primers, PCR amplification was performed on the genomic DNAs of 36 blood samples (3 goat species and 3 sheep species), with the PCR product obtained using F1-1/R1-1 as the primer being clear and reproducible. F1-1/R1-1 was used as the primer in the amplification of the COI fragment of chicken, pork, donkey meat, beef, Laiwu black goat meat, mutton sheep meat and duck meat samples, and then the amplified fragments of the seven kinds of meat samples were sequenced. The sequence homology between goat and sheep was only 36.62%, and the sequence difference among other breeds was also obvious The results showed that F1-1/R1-1 can be used as a special primer to distinguish effectively between mutton meat and meat from other animal sources, and also between goat meat and sheep meat samples.