食品科学 ›› 2009, Vol. 30 ›› Issue (19): 185-189.doi: 10.7506/spkx1002-6630-200919041

• 生物工程 • 上一篇    下一篇

检测黄曲霉毒素生物合成相关基因的芯片的研制

胡娜,许洋,涂追   

  1. 1.江西科技师范学院生命科学院 2.南昌大学中德联合研究院
  • 收稿日期:2008-09-22 修回日期:2009-03-26 出版日期:2009-10-01 发布日期:2010-12-29
  • 通讯作者: 许杨 E-mail:na9728067@yahoo.com.cn
  • 基金资助:

    江西科技师范学院青年创新基金项目

Development of Gene Chip for Aflatoxin Detection

HU Na1,XU Yang2,*,TU Zhui2   

  1. 1. College of Life Science, Jiangxi Science and Technology Normal University, Nanchang 330013, China ;
    2. Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, China
  • Received:2008-09-22 Revised:2009-03-26 Online:2009-10-01 Published:2010-12-29
  • Contact: XU Yang E-mail:na9728067@yahoo.com.cn

摘要:

本试验建立了比较成熟的可用于分析黄曲霉毒素生物合成相关基因的芯片技术。研究表明芯片点阵后依次通过温浴2h,650 mJ/cm2紫外交联30s,80℃烘烤2h,预杂交45 min,清洗,干燥等步骤,最后与待测样品在42℃杂交16h,可获得低背景高质量的芯片。本实验所用探针来源于寄生曲霉,而待测样品选择了黄曲霉菌株,芯片扫描结果显示荧光信号稳定,与反转录PCR扩增结果一致,并且无背景干扰。证明探针设计合理,实验方法可靠。初步建立了用芯片检测与黄曲霉毒素生物合成相关基因的技术平台。

关键词: 寄生曲霉, 黄曲霉, 黄曲霉毒素, 基因芯片

Abstract:

In order to detect genes related to aflatoxin biosynthesis, a high quality gene chip was successfully developed here. The preparation of this gene chip was involved in a series of processes: array on the slide, heating in 42 water bath for 2 h, UV-induced crosslink at 650 mJ/cm2 for 30 seconds, baking in oven at 80 for 2 h, pre-hybridization for 45 min, then washing and drying, and final hybridization with fluor-derivatized samples at 42 for 16 h. The probe originated from Aspergillus parasiticus could be used to analyze aflatoxin gene expression in Aspergillus flavus. Therefore, this probe can provide stable fluorescence signal without background inference and results consistent with reverse transcriptase polymerase chain reaction (RT-PCR).

Key words: Aspergillus parasiticus, Aspergillus flavus, aflatoxin, gene chip

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