Improvement of Methods for Safety Control of Microbial Producers and Food Produced Therewith.
DNA extraction
DNA sequencing
PCR assay
genetically modified microorganisms
microbial producers
Journal
Bulletin of experimental biology and medicine
ISSN: 1573-8221
Titre abrégé: Bull Exp Biol Med
Pays: United States
ID NLM: 0372557
Informations de publication
Date de publication:
Jul 2023
Jul 2023
Historique:
received:
29
12
2022
medline:
22
8
2023
pubmed:
10
8
2023
entrez:
10
8
2023
Statut:
ppublish
Résumé
The study substantiates the necessity to implement the algorithm of molecular-genetic assessment of biosafety of the genetically modified microorganisms (GMM) and to develop standardized methods to test the genetically modified strains producing enzymes, bioactive substances, and other products of microbial synthesis prior to their use in food industry. Analysis of microbial producers and related food products for the presence of GMM-associated DNA revealed high incidence of the marker genes amp and lacZ in enzyme preparations and in mycelium of industrial genetically modified producer of Aspergillus genus. The procedure of extraction of DNA from mycelium of mold fungi is optimized by including the stage of additional purification of the extracts, assessment of their purity by PCR with universal ITS primers, and determination of effective DNA concentration in the samples prior to conduction of the molecular genetic assay. For identification and genotyping of mold fungi (the biotechnological producers of enzyme preparations), the Sanger sequencing method was adapted. Using this modified method, we determined the species of five equivocally identified strains of Aspergillus genus. To identify the closely-related micromycetes of Ascomycota division, a genotyping algorithm was developed based on amplification of total DNA with expanded panel of primers and DNA sequencing by capillary electrophoresis.
Identifiants
pubmed: 37561374
doi: 10.1007/s10517-023-05874-3
pii: 10.1007/s10517-023-05874-3
doi:
Substances chimiques
DNA
9007-49-2
DNA Primers
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
393-398Informations de copyright
© 2023. Springer Science+Business Media, LLC, part of Springer Nature.
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