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Acta Agron Sin ›› 2009, Vol. 35 ›› Issue (8): 1432-1438.doi: 10.3724/SP.J.1006.2009.01432

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Developing a Method of Oligonucleotide Microarray for Event Specific Detection of Transgenic Maize(Zea mays

LU Xing-Bo1,WU Hai-Bin12,WANG Min2,LI Bao-Du2,YANG Chong-Liang1,SUN Hong-Wei1*   

  1. 1Plant Protection Research Institute,Shandong Academy of Agricultural Sciences,Jinan 250100,China;2College of Plant Protection,Qingdao Agricultural University,Qingdao 266109,China
  • Received:2008-12-08 Revised:2009-03-20 Online:2009-08-12 Published:2009-06-10
  • Contact: SUN Hong-Wei, E-mail: hongweisun@126.com

Abstract:

With the spread of genetically modified organism (GMO), the food and environment securities related to GMO attract great attention to the public. Genetically modified maize (Zea mays L.) is one of the most popular GMO varieties, which accounts for 22.5% of total planting area of transgenic crops. Currently, more than 40 countries and regions govern GMO food products with a compulsory tagging policy. Thus, technique for testing transgenic plants is of great importance, such as screening, gene specific, construct specific, event specific detection method, which have become a mainstay of GMOs detection. Event specific microarray shows a prosperous application due to its advantages of its high specificity, high sensitivity, automation and high efficiency. In this study, on the base of integrated DNA constructs of seven genetically modified maize varieties, Bt11, Bt176, Mon810, Mon863, TC1507, GA21, and NK603, event specific primers were designed for polymerase chain reaction (PCR) using Primer premier 5.0, and the PCR system and program were optimized. The results indicated that the sensitivity of PCR detection was 0.1%. In addition, based on unique and specific integration junction sequences between the host plant genome DNA and the integrated gene, a 40mer event specific oligo probe with high specificity was designed for each variety using Primer premier 5.0 and Oligo 6.0, all the event specific probes were developed in one microarray. To ensure the reliability of microarray detection, we used different kinds of positive and negative controls in oligonucleotide microarray. The results indicate that the sensitivity of microarray detection (0.01%) and the event specific oligonucleotide probes developed in this study meet the requirements of sensitivity and specificity for detecting and identifying genetically modified maize, which enhances the testing accuracy and efficiency.

Key words: Transgenic maize, Event specific, Oligonucletide microarray, Oligo probe, Sensitivity

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