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Integrated DNA walking system to characterize a broad spectrum of GMOs in food/feed matrices

Marie-Alice Fraiture, Philippe Herman, Loic Lefèvre, Isabel Taverniers, Marc De Loose UGent, Dieter Deforce UGent and Nancy H Roosens (2015) BMC BIOTECHNOLOGY. 15.
abstract
Background: In order to provide a system fully integrated with qPCR screening, usually used in GMO routine analysis, as well as being able to detect, characterize and identify a broad spectrum of GMOs in food/feed matrices, two bidirectional DNA walking methods targeting p35S or tNOS, the most common transgenic elements found in GM crops, were developed. These newly developed DNA walking methods are completing the previously implemented DNA walking method targeting the t35S pCAMBIA element. Results: First, the newly developed DNA walking methods, anchored on the sequences used for the p35S or tNOS qPCR screening, were tested on Bt rice that contains these two transgenic elements. Second, the methods were assessed on a maize sample containing a low amount of the GM MON863 event, representing a more complex matrix in terms of genome size and sensitivity. Finally, to illustrate its applicability in GMO routine analysis by enforcement laboratories, the entire workflow of the integrated strategy, including qPCR screening to detect the potential presence of GMOs and the subsequent DNA walking methods to characterize and identify the detected GMOs, was applied on a GeMMA Scheme Proficiency Test matrix. Via the characterization of the transgene flanking region between the transgenic cassette and the plant genome as well as of a part of the transgenic cassette, the presence of GMOs was properly confirmed or infirmed in all tested samples. Conclusion: Due to their simple procedure and their short time-frame to get results, the developed DNA walking methods proposed here can be easily implemented in GMO routine analysis by the enforcement laboratories. In providing crucial information about the transgene flanking regions and/or the transgenic cassettes, this DNA walking strategy is a key molecular tool to prove the presence of GMOs in any given food/feed matrix.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
REAL-TIME PCR, FLANKING SEQUENCE DETERMINATION, UNAUTHORIZED GMOS, PRODUCTS, GENES, FOOD
journal title
BMC BIOTECHNOLOGY
BMC Biotechnol.
volume
15
article number
76
pages
11 pages
Web of Science type
Article
Web of Science id
000359686500001
JCR category
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
JCR impact factor
2.452 (2015)
JCR rank
63/161 (2015)
JCR quartile
2 (2015)
ISSN
1472-6750
DOI
10.1186/s12896-015-0191-3
language
English
UGent publication?
yes
classification
A1
copyright statement
I have retained and own the full copyright for this publication
id
6842118
handle
http://hdl.handle.net/1854/LU-6842118
date created
2015-06-23 08:49:03
date last changed
2016-12-21 15:42:03
@article{6842118,
  abstract     = {Background: In order to provide a system fully integrated with qPCR screening, usually used in GMO routine analysis, as well as being able to detect, characterize and identify a broad spectrum of GMOs in food/feed matrices, two bidirectional DNA walking methods targeting p35S or tNOS, the most common transgenic elements found in GM crops, were developed. These newly developed DNA walking methods are completing the previously implemented DNA walking method targeting the t35S pCAMBIA element.
Results: First, the newly developed DNA walking methods, anchored on the sequences used for the p35S or tNOS qPCR screening, were tested on Bt rice that contains these two transgenic elements. Second, the methods were assessed on a maize sample containing a low amount of the GM MON863 event, representing a more complex matrix in terms of genome size and sensitivity. Finally, to illustrate its applicability in GMO routine analysis by enforcement laboratories, the entire workflow of the integrated strategy, including qPCR screening to detect the potential presence of GMOs and the subsequent DNA walking methods to characterize and identify the detected GMOs, was applied on a GeMMA Scheme Proficiency Test matrix. Via the characterization of the transgene flanking region between the transgenic cassette and the plant genome as well as of a part of the transgenic cassette, the presence of GMOs was properly confirmed or infirmed in all tested samples.
Conclusion: Due to their simple procedure and their short time-frame to get results, the developed DNA walking methods proposed here can be easily implemented in GMO routine analysis by the enforcement laboratories. In providing crucial information about the transgene flanking regions and/or the transgenic cassettes, this DNA walking strategy is a key molecular tool to prove the presence of GMOs in any given food/feed matrix.},
  articleno    = {76},
  author       = {Fraiture, Marie-Alice and Herman, Philippe and Lef{\`e}vre, Loic and Taverniers, Isabel and De Loose, Marc and Deforce, Dieter and Roosens, Nancy H},
  issn         = {1472-6750},
  journal      = {BMC BIOTECHNOLOGY},
  keyword      = {REAL-TIME PCR,FLANKING SEQUENCE DETERMINATION,UNAUTHORIZED GMOS,PRODUCTS,GENES,FOOD},
  language     = {eng},
  pages        = {11},
  title        = {Integrated DNA walking system to characterize a broad spectrum of GMOs in food/feed matrices},
  url          = {http://dx.doi.org/10.1186/s12896-015-0191-3},
  volume       = {15},
  year         = {2015},
}

Chicago
Fraiture, Marie-Alice, Philippe Herman, Loic Lefèvre, Isabel Taverniers, Marc De Loose, Dieter Deforce, and Nancy H Roosens. 2015. “Integrated DNA Walking System to Characterize a Broad Spectrum of GMOs in Food/feed Matrices.” Bmc Biotechnology 15.
APA
Fraiture, M.-A., Herman, P., Lefèvre, L., Taverniers, I., De Loose, M., Deforce, D., & Roosens, N. H. (2015). Integrated DNA walking system to characterize a broad spectrum of GMOs in food/feed matrices. BMC BIOTECHNOLOGY, 15.
Vancouver
1.
Fraiture M-A, Herman P, Lefèvre L, Taverniers I, De Loose M, Deforce D, et al. Integrated DNA walking system to characterize a broad spectrum of GMOs in food/feed matrices. BMC BIOTECHNOLOGY. 2015;15.
MLA
Fraiture, Marie-Alice, Philippe Herman, Loic Lefèvre, et al. “Integrated DNA Walking System to Characterize a Broad Spectrum of GMOs in Food/feed Matrices.” BMC BIOTECHNOLOGY 15 (2015): n. pag. Print.