Development of a real-time PCR marker targeting a new unauthorized genetically modified microorganism producing protease identified by DNA walking.
Antimicrobial resistance genes
DNA walking anchored on pUB110
Food and feed safety
Genetically modified bacteria
Protease
Real-time PCR detection
Journal
International journal of food microbiology
ISSN: 1879-3460
Titre abrégé: Int J Food Microbiol
Pays: Netherlands
ID NLM: 8412849
Informations de publication
Date de publication:
16 Sep 2021
16 Sep 2021
Historique:
received:
08
04
2021
revised:
13
07
2021
accepted:
15
07
2021
pubmed:
26
7
2021
medline:
25
9
2021
entrez:
25
7
2021
Statut:
ppublish
Résumé
A PCR-based DNA walking analysis was performed on a protease product suspected to contain a new unauthorized genetically modified microorganism (GMM). Though the characterization of unnatural associations of sequences between the pUB110 shuttle vector and a Bacillus amyloliquefaciens gene coding for a protease, the presence of the GMM was shown. Based on these sequences of interest, a real-time PCR marker was developed to target specifically the newly discovered GMM, namely GMM protease2. The performance of the real-time PCR marker was assessed in terms of specificity and sensitivity. The applicability of the real-time PCR GMM protease2 marker was also demonstrated on microbial fermentation products. To confirm its use by other GMO enforcement laboratories, the transferability of the in-house validated real-time PCR marker was demonstrated by assays performed by an external laboratory.
Identifiants
pubmed: 34303961
pii: S0168-1605(21)00289-0
doi: 10.1016/j.ijfoodmicro.2021.109330
pii:
doi:
Substances chimiques
Genetic Markers
0
Peptide Hydrolases
EC 3.4.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
109330Informations de copyright
Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.