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运用基因芯片技术检测牛、山羊、猪和鸡源性成分
引用本文:石丰运,缪建锟,张利平,陶虹,吕建强,阮周曦,宗卉. 运用基因芯片技术检测牛、山羊、猪和鸡源性成分[J]. 生物工程学报, 2010, 26(6): 823-829
作者姓名:石丰运  缪建锟  张利平  陶虹  吕建强  阮周曦  宗卉
作者单位:1. 甘肃农业大学动物科学技术学院,兰州730070;深圳出入境检验检疫局,深圳518010
2. 中国农业大学农业生物技术国家重点实验室,北京,100193
3. 甘肃农业大学动物科学技术学院,兰州,730070
4. 深圳出入境检验检疫局,深圳,518010
基金项目:深圳出入境检验检疫局科研项目 (No. SZ2008015) 资助。
摘    要:本研究通过对脊椎动物分子标记基因进行序列分析,最终选择线粒体DNA(mtDNA)16S rRNA基因为目标基因,利用一对通用引物,在该引物扩增区间设计了4条特异性基因芯片检测探针及2条质控探针用于对牛、山羊、猪、鸡等4种动物源性成分进行检测。通过对PCR扩增体系及杂交体系的优化,该检测方法能实现对上述4种动物源性成分同时进行快速、准确地检测,具有很好的特异性,灵敏度均达到1pg,最终建立了这4种动物源性基因芯片检测方法。该基因芯片检测技术将为我国进出口饲料中的动物源性成分的鉴别提供新的检测方法和技术支持。

关 键 词:线粒体16S rRNA基因,DNA芯片,动物源性
收稿时间:2010-01-14

Detection of bovine, goat, pig and chicken derived ingredients in animal products with universal PCR-microarray method
Fengyun Shi,Jiankun Miao,Liping Zhang,Hong Tao,Jianqiang Lü,Zhouxi Ruan,Hui Zong. Detection of bovine, goat, pig and chicken derived ingredients in animal products with universal PCR-microarray method[J]. Chinese journal of biotechnology, 2010, 26(6): 823-829
Authors:Fengyun Shi  Jiankun Miao  Liping Zhang  Hong Tao  Jianqiang Lü  Zhouxi Ruan  Hui Zong
Affiliation:Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China; Shenzhen Entry-exit Inspection and Quarantine Bureau, Shenzhen 518010, China;State Key Laboratory of Agro-biotechnology, China Agricultural University, Beijing 100193, China;Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;Shenzhen Entry-exit Inspection and Quarantine Bureau, Shenzhen 518010, China;Shenzhen Entry-exit Inspection and Quarantine Bureau, Shenzhen 518010, China;Shenzhen Entry-exit Inspection and Quarantine Bureau, Shenzhen 518010, China;Shenzhen Entry-exit Inspection and Quarantine Bureau, Shenzhen 518010, China
Abstract:We analyzed the sequence of vertebrate molecular marker genes, then we selected the mitochondrial DNA (mtDNA) 16S rRNA gene as marker gene. In order to detect four kinds of animal-derived ingredients, which including bovine, goat, pig and chicken. We utilized a pair of universal primers, designed four sets of species-specific microarray probes and two pairs of quality control probes. We optimized the PCR amplifications and hybridization conditions, therefore these four kinds of animal-derived ingredients could be rapid and accurate detected by this approach. The detection limits were all reaches 1 pg. We established the detection platform of these four kinds of animal-derived ingredients. This universal PCR-microarray assay provides a new method for the identification of animal-derived ingredients in the import-export field.
Keywords:mitochondrion 16S rRNA gene   DNA chip   animal-derived ingredients
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