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Development of an event-specific Real-time PCR detection method for the transgenic Bt rice line KMD1

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Abstract

The present study describes an event-specific quantitative Real-time PCR detection method for the transgenic Bt rice line Kemingdao 1 (KMD1). This rice line which is not approved in any country so far is likely to be approved in China in the near future. The developed primers amplify a DNA sequence spanning the integration site of the genetic construct in KMD1. DNA sequence information of this unknown site necessary for primer design was achieved using SiteFinding-PCR technique. The specificity of the detection method was shown by testing a number of different transgenic and conventional plant varieties (e.g. rice LL 601, LL 62, Bt 63). As alternative to genomic DNA, we developed double target hybrid amplicons as synthetic calibration standards in Real-time PCR. These amplicons contained both one copy of the KMD1 event-specific sequence and one copy of a sequence of the rice reference gene gos9. The limit of quantification (LOQ) of the method was tested to be 0.05%.

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Acknowledgments

This work was funded by the Bavarian State Ministry of the Environment, Public Health and Consumer Protection.

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Authors and Affiliations

  1. Lehrstuhl für Allgemeine Lebensmitteltechnologie, Technische Universität München, Am Forum 2, 85350, Freising-Weihenstephan, Germany

    Ruzha Babekova & Karl-Heinz Engel

  2. Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Veterinaerstr. 2, 85764, Oberschleissheim, Germany

    Tristan Funk, Sven Pecoraro & Ulrich Busch

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  1. Ruzha Babekova
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  2. Tristan Funk
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  3. Sven Pecoraro
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  4. Karl-Heinz Engel
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  5. Ulrich Busch
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Correspondence to Sven Pecoraro.

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R. Babekova and T. Funk contributed equally to this paper.

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Babekova, R., Funk, T., Pecoraro, S. et al. Development of an event-specific Real-time PCR detection method for the transgenic Bt rice line KMD1. Eur Food Res Technol 228, 707–716 (2009). https://doi.org/10.1007/s00217-008-0981-0

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  • DOI: https://doi.org/10.1007/s00217-008-0981-0

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