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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渤海地区鱼种鉴定

李潇1, 曲焱焱1, 张丕乔2, 张津3, 张丽华1*, 黄大亮2, 张玉奎1   

  1. 1. 中国科学院大连化学物理研究所, 中国科学院分离分析化学重点实验室, 辽宁 大连 116023; 2. 庄河出入境检验检疫局, 辽宁 庄河 121013; 3. 安捷伦科技(中国)有限公司, 北京 100102
  • 收稿日期:2011-03-31 修回日期:2011-05-24 出版日期:2011-07-28 发布日期:2011-08-19
  • 通讯作者: 张丽华,博士,研究员,博士生导师,主要从事复杂体系的分离鉴定. Tel: (0411)84379720
  • 基金资助:

    国家科技支撑计划项目（2009BAK59B02）

Application of polymerase chain reaction-restriction fragment length polymorphism and lab-on-a-chip technology to the identification of fish species from Bohai Bay

LI Xiao1, QU Yanyan1, ZHANG Piqiao2, ZHANG Jin3, ZHANG Lihua1*, HUANG Daliang2, ZHANG Yukui1   

  1. 1. Dalian Institute of Chemical Physics, CAS Key Laboratory of Separation Science for Analytical Chemistry, Chinese Academy of Sciences, Dalian 116023, China; 2. Zhuanghe Entry-Exit Inspection and Quarantine Bureau, Zhuanghe 121013, China; 3. Agilent Technologies Co., Ltd. (China), Beijing 100102, China
  • Received:2011-03-31 Revised:2011-05-24 Online:2011-07-28 Published:2011-08-19

PDF

420

被引次数

6

摘要： 以渤海湾具有重要商业价值的9种海鱼(小黄鱼、花鲈、蓝点马鲛、鲐、钝吻黄盖鲽、棘头梅童鱼、许氏平鲉、高眼鲽和长蛇鲻)为研究对象,利用聚合酶链式反应(PCR)-限制性片段长度多态性(RFLP)和芯片生物分析技术对其进行鱼种鉴定。首先提取其基因组脱氧核糖核酸(DNA),对其细胞色素b的特定片段(464 bp)进行扩增,然后选择DdeI、HaeIII和NlaIII3种限制性内切酶进行酶切,并利用芯片生物分析技术得到酶切产物的特异图谱和确切的片段大小,从而有效区分了9种海鱼。研究结果表明,PCR-RFLP和芯片生物分析技术在鱼种鉴定上具有精确、鉴别和快速三大优势,可为鱼类食品检测提供科学依据。

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

Abstract: Nine representative fish species from Bohai Bay were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and lab-on-a-chip technology. The nine fish species were Pseudosciaena polyactis, Lateolabrax japonicus, Scomberomorus niphonius, Pneumatophorus japonicus, Pseudopleuronectes yokohamae, Collichthys lucidus, Sebastes schlegeli, Cleisthenes herzensteini and Saurida elongata. The 464 bp fragment of mitochondrial cytochrome b gene was amplified and further digested by DdeI, HaeIII and NlaIII. The digested DNA fragments were analyzed by microfluidic capillary electrophoresis. The nine fish species were fully discriminated. The results demonstrated that the proposed method is efficient, precise and fast in fish species identification.

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