摘要
依据牛１．７０９卫星ＤＮＡ序列设计了１对引物，建立了ＰＣＲ鉴定生、熟牛肉的方法。应用本方法特异地扩增出预期的牛２１８ｂｐＤＮＡ片段，其检测敏感度对生牛肉达３３．６ｆｇＤＮＡ，对熟牛肉和高压牛肉为０．３２ｐｇ。运用该引物均可扩增出水牛、牦牛、奶牛、黄牛肉单一的相同大小的ＤＮＡ条带，而对马、山羊、绵羊、骆驼、鹿、猪等１５种动物肉的ＤＮＡ扩增则呈阴性。扩增片段经ＨａｅⅢ酶切分析确认，所得１２９、７９、１０ｂｐ片段与微机分析结果一致。利用本法对１０３份生、熟牛肉及其制品进行鉴定，检出率为１００％。对各种样品检测，均可在６ｈ内完成。
Genomic DNA extracted from beef samples was amplified by the poly-merase chain reaction for identification of beef.The sequence selected for amplificationconsisted of 218 bp lying in the 1.709 satellite DNA of bovine,a pair of syntheticoligonucleotides flanking this sequence were used as primers. The amplified productswere subjected to rapid electrophoresis in an agarose gel and visualized under ultravioletillumination after EB staining. A Hae Ⅲ restriction endonuclease test was simultaneous-ly made for verifying the specificity of the PCR amplification. The results showed that aspecific amplification fragment of the expected size was detected and demonstrated thatit was positive for bovine,cow,buffalo and yak meat,but negative for equine,ovine,goat,camel,deer,swine and mouse meat etc. An amount as small as 33.6 fg totalDNA from raw beef samples and 0.324 pg DNA from cooked or autoclaved beef samples could be detected by PCR.
出处
《中国兽医学报》
CAS
CSCD
1996年第2期179-183,共5页
Chinese Journal of Veterinary Science
关键词
聚合酶链反应
牛肉
鉴定
identification
beef
polymerase chain reaction
