研究论文

曲霉属产毒真菌DNA的制备及其毒素相关基因aflR的PCR 检测

  • 蒋丹 ,
  • 靳卫林 ,
  • 苏德山 ,
  • 黄耀江
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  • 1. 中央民族大学, 北京 100081;
    2. 北京市食品环境与健康工程技术研究中心, 北京 100081;
    3. 辽宁出入境检验检疫局检验检疫技术中心, 大连 116001
蒋丹,副研究员,研究方向为食品安全与健康,电子信箱:jiangdan66@163.com;靳卫林(共同第一作者),硕士研究生,研究方向为分子生态学,电子信箱:jwldsg@126.com

收稿日期: 2014-01-17

  修回日期: 2014-03-26

  网络出版日期: 2014-05-29

基金资助

教育部新世纪优秀人才项目(NCET-11-0842);北京市科技创新基地培育与发展工程项目(Z131106002813027)

DNA Preparation of Toxigenic Aspergillus and PCR Detection of aflR Toxin Gene

  • JIANG Dan ,
  • JIN Weilin ,
  • SU Deshan ,
  • HUANG Yaojiang
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  • 1. Minzu University of China, Beijing 100081, China;
    2. Beijing Engineering Research Center of Food Environment and Public Health, Beijing 100081, China;
    3. Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China

Received date: 2014-01-17

  Revised date: 2014-03-26

  Online published: 2014-05-29

摘要

研究了用改良的CTAB 法提取曲霉属产毒真菌DNA 的方法,并与试剂盒提取法进行比较。经过改良的CTAB 法能够成功提取出曲霉属产毒真菌的DNA,且浓度和纯度均可达到PCR 扩增的标准。根据合成黄曲霉毒素的aflR 基因设计两对引物,运用PCR 方法进行扩增鉴定,结果表明黄曲霉、寄生曲霉和溜曲霉基因中可检出aflR 基因,烟曲霉和杂色曲霉中并未检出,达到对黄曲霉毒素产生菌进行鉴定的目的。这种方法可为产毒真菌的快速检测提供参考。

本文引用格式

蒋丹 , 靳卫林 , 苏德山 , 黄耀江 . 曲霉属产毒真菌DNA的制备及其毒素相关基因aflR的PCR 检测[J]. 科技导报, 2014 , 32(14) : 31 -34 . DOI: 10.3981/j.issn.1000-7857.2014.14.004

Abstract

This paper discusses the modified CTAB method for DNA extraction of toxigenic Aspergillus and compares it with the kit extraction method. The modified CTAB method successfully extracted DNA of toxigenic Aspergillus. The concentration and purity reached the standard of PCR amplification. Two pairs of primers were designed based on the aflR gene for aflatoxin biosynthesis regulation, and the toxigenic Aspergillus aflR gene was identified by using PCR amplification. The results show that the aflR gene is contained in the genome of Aspergillus flavus, Aspergillus parasitic and Aspergillus tamarii, Aspergillus fumigates and Aspergillus versicolor are not detected, realizing the purpose for identifying aflatoxin producing strains. This method provides a reference for rapid detection of toxigenic fungi.

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