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鹌鹑源性成分PCR检测方法的建立
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张慧霞1,张利平1,吴建平1,阮周曦2,宗卉2*
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（1.甘肃农业大学 动物科学技术学院,甘肃 兰州730070;
2.进出口深圳检验检疫局 动物与植物检验检疫技术中心,广东 深圳518010)
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摘要：
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根据GenBank中的鹌鹑线粒体DNA(mtDNA) 12sRNA基因保守序列，用Primer5.0软件设计了1对针对鹌鹑的特异性扩增引物，建立了一种检测鹌鹑动物源性成分的PCR 方法。通过聚合酶链式反应(PCR）从鹌鹑肉和鹌鹑肉骨粉样品中扩增到了235bp的目的片段，而参考物种鸽、鸡、鸭、鹅、鸵鸟、鹧鸪、牛、羊、马、驴、鱼和空白对照则无目的片段，说明该引物只对鹌鹑有特异性。测序结果表明，鹌鹑组织PCR产物的核苷酸序列与GenBank中检索到的相应序列相吻合；该方法的检测灵敏度为1g/kg。 |
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关键词：
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鹌鹑；线粒体DNA；动物源性成分；聚合酶链式反应 |
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中图分类号：S 851.347.2：Q 503 文献标识码：
A 文章编号：1673-4696(2008)04-0346-04 |
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Establishment of PCR assay for detection of quail materials |
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ZHANG Hui-xia1,ZHANG Li-ping1,WU
Jian-ping1,RUAN Zhou-xi2,ZONG Hui2 |
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(1.College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;2.Technology
Center of Inspection and Quarantine for Animal and Plant,Shenzhen Inspection and Quarantine Bureau,
Shenzhen 518010,China) |
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Abstract: Species-specific primers were designed by Primer 5.0 software according to the conserved sequence of quail mitochondrial DNA(mtDNA) 12sRNA gene available in GenBank.The 235bp fragments were amplified from quail and meat and bone meals of quail by PCR.But no fragments were amplified from pigeon,chicken,duck,goose,ostrich,partridge,cow,sheep,horse,donkey,fish and blank control.The specifiction of the PCR was demonstrated by direct sequencing of the amplified products from animal feedstuff.The results of DNA sequencing agreed with sequences of quail in GenBank.The method was sensitive to quail DNA in derived materials with a detection limit of 1g/kg,so it could become an effective and precise method for quail-derived materials. |
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Key words: quail；mitochondrial DNA（mtDNA）;animal materials;PCR |
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