色谱

色谱

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聚合酶链式反应-限制性片段长度多态性和芯片生物分析技术用于台湾海峡常见石斑鱼和鲷鱼的品种鉴定

陈双雅1, 张津2, 陈伟玲1, 徐敦明1, 周昱1*   

  1. 1. 厦门出入境检验检疫局检验检疫技术中心, 福建 厦门 361026; 2. 安捷伦科技（中国）有限公司, 北京 100102
  • 收稿日期:2011-04-20 修回日期:2011-04-29 出版日期:2011-07-28 发布日期:2011-08-19
  • 通讯作者: 周昱,研究员,主要研究方向为食品安全. Tel: (0592)3269929
  • 基金资助:

    国家质检总局科技项目(No. 2011IK245)

Species identification of grouper and snapper in Taiwan Strait using polymerase chain reaction-restriction fragment length polymorphism analysis and lab-on-a-chip system

CHEN Shuangya1, ZHANG Jin2, CHEN Weiling1, XU Dunming1, ZHOU Yu1*   

  1. 1. Inspection and Quarantine Technology Center, Xiamen Entry-Exit Inspection and Quarantine Bureau, Xiamen 361026, China; 2. Agilent Technologies Co., Ltd.(China), Beijing 100102, China
  • Received:2011-04-20 Revised:2011-04-29 Online:2011-07-28 Published:2011-08-19

PDF

458

被引次数

6

摘要： 应用聚合酶链式反应-限制性片段长度多态性分析(PCR-RFLP)和芯片生物分析系统建立了台湾海峡常见石斑鱼和鲷鱼的分子生物学品种鉴定新方法。首先提取鱼的基因组脱氧核糖核酸(DNA)进行细胞色素b基因特定片段的PCR扩增,然后用DdeI、HaeIII和NlaIII 3种限制性内切酶进行酶切,在Agilent DNA 1000芯片上对酶切片段进行分离。该方法成功鉴定了台湾海峡常见的8种石斑鱼品种和5种鲷鱼品种,是一种快速、简便、有效的鱼类品种鉴定分析手段。

关键词: 鲷鱼, 聚合酶链式反应-限制性片段长度多态分析, 石斑鱼, 物种鉴定, 细胞色素b, 芯片实验室

Abstract: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and lab-on-a-chip system were used to identify grouper and snapper species in Taiwan Strait. A fragment of 464 bp length of mitochondrial cytochrome b gene was amplified by PCR and the products were digested with restriction enzymes DdeI, HaeIII and NlaIII, individually. The fragments generated after digestion were further resolved on the DNA Chip. Eight grouper species and five snapper species were successfully identified. The results demonstrated that PCR-RFLP analysis and lab-on-a-chip system provide a fast, easy, automated, and reliable analysis approach. This approach is potential for the purpose of fish adulteration control.

Key words: cytochrome b, grouper, snapper, species identification, lab-on-a-chip, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)