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Construction of Standard Reference Molecule and Its Application in Event-specific Transgenic Detection of Genetically Modified Maize 59122

LU Jiao，YANG Rong，HUANG Kun-lun，ZHAO Wei-wei，LUO Yun-bo，YUAN Yan-fang，XU Wen-tao   

  1. 1. Food Safety Laboratory, College of Food Science and Nutritional Engineering, China Agricultural University,
    Beijing 100083, China ；2. Supervision, Inspection and Testing Center of Genetically
    Modified Organisms (Beijing), Ministry of Agriculture, 100083 Beijing, China
  • Online:2011-01-25 Published:2011-01-10
  • Contact: XU Wen-tao

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Abstract: Transgenic event-specific primers and Taqman probes based on the left-flanking sequence of 59122 were
designed, and a real-time PCR method was developed to quantitatively detect the event-specific genetically modified maize
59122. The standard reference molecule (integration of the reference gene sequence and the left-wing sequence) with a
gradient range of 101－105 and a quantitative range of 0.01%－100% were obtained by a recombinant PCR method. Two
standard curves of the reference gene sequence and the left-wing sequence were established with acceptable CV (coefficient
of variance), SD (standard deviation) and R2 (correlation coefficient). Five mixed samples with known genetically modified
contents of 0.1%, 0.5%, 1%, 2% and 5% were determined with the results of 0.12%, 0.56%, 1.04%, 1.61% and 4.66%,
respectively. The experimental error was less than 25%. The assay can be applied to analyze genetically modified content (raw
materials as well as processed food) and will provide a technical support for the preparation of standard reference molecule.

Key words: event-specific, real-time PCR, genetically modified maize, standard reference molecule

CLC Number: