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BMC Biotechnol. 2018 Jun 27;18(1):40. doi: 10.1186/s12896-018-0446-x.

Development and validation of an integrated DNA walking strategy to detect GMO expressing cry genes.

Author information

1
Scientific Institute of Public Health (WIV-ISP), Platform of Biotechnology and Bioinformatics (PBB) by: Sciensano, Transversal & Applied Genomics (TAG), J. Wytsmanstraat 14, 1050, Brussels, Belgium.
2
Scientific Institute of Public Health (WIV-ISP), Operational Direction Expertise, Service provisions & Customer relations by: Sciensano, Scientific Direction Expertise, Service provisions & Customer relations, J. Wytsmanstraat 14, 1050, Brussels, Belgium.
3
Scientific Institute of Public Health (WIV-ISP), Platform of Biotechnology and Bioinformatics (PBB) by: Sciensano, Transversal & Applied Genomics (TAG), J. Wytsmanstraat 14, 1050, Brussels, Belgium. nancy.roosens@wiv-isp.be.

Abstract

BACKGROUND:

Recently, an integrated DNA walking strategy has been proposed to prove the presence of GMO via the characterisation of sequences of interest, including their transgene flanking regions and the unnatural associations of elements in their transgenic cassettes. To this end, the p35S, tNOS and t35S pCAMBIA elements have been selected as key targets, allowing the coverage of most of GMO, EU authorized or not. In the present study, a bidirectional DNA walking method anchored on the CryAb/c genes is proposed with the aim to cover additional GMO and additional sequences of interest.

RESULTS:

The performance of the proposed bidirectional DNA walking method anchored on the CryAb/c genes has been evaluated in a first time for its feasibility using several GM events possessing these CryAb/c genes. Afterwards, its sensitivity has been investigated through low concentrations of targets (as low as 20 HGE). In addition, to illustrate its applicability, the entire workflow has been tested on a sample mimicking food/feed matrices analysed in GMO routine analysis.

CONCLUSION:

Given the successful assessment of its performance, the present bidirectional DNA walking method anchored on the CryAb/c genes can easily be implemented in GMO routine analysis by the enforcement laboratories and allows completing the entire DNA walking strategy in targeting an additional transgenic element frequently found in GMO.

KEYWORDS:

Characterization; DNA walking; Detection; GMO; Identification; qPCR

PMID:
29945581
PMCID:
PMC6020286
DOI:
10.1186/s12896-018-0446-x
[Indexed for MEDLINE]
Free PMC Article

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