Use of cloned DNA fragments for event-specific quantification of genetically modified organisms in pure and mixed food products

Isabel Taverniers, Pieter Windels, Erik Van Bockstaele, Marc De Loose

Onderzoeksoutput: Bijdrage aan tijdschriftA1: Web of Science-artikel


An event-specific PCR method for detection and
quantification of genetically modified Roundup Ready soybean
(RRS) is described in this article. The complete DNA
sequence at both the right and left integration sites of this
genetically modified organism has recently been determined.
Based on these sequence data, transformation eventspecific
primer pairs were developed. These primers amplify
a fragment of the unique junction region between the inserted
DNA and the plant DNA and therefore act as unique
identifiers. Two sensitive, qualitative PCR assays gave absolute
detection limits of 5 copies of the RRS junction fragment
in 100 pg of total DNA per reaction. A real-time PCR
method was then developed with the LightCycler System.
For determination of the RRS content, a completely new
type of external calibration standard is introduced here. A
fragment of the RRS specific junction region and a fragment
of the endogenous soybean lectin gene were both
cloned in a plasmid vector. These new diagnostic DNA
fragments allow quantification of RRS in whichever type of
matrix, in a range of 10–106 copies of each target.
TijdschriftEuropean Food Research and Technology
Pagina's (van-tot)417-424
Aantal pagina's8
StatusGepubliceerd - 2001

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