Quantification of the 35S Promoter in DNA Extracts from Genetically Modified Organisms Using Real-Time Polymerase Chain Reaction and Specificity Assessment on Various Genetically Modified Organisms, Part I: Operating Procedure
| Autor: | Cécile Collonnier, Max Feinberg, Francine Boyer, Sophie Fernandez, Georges Berthier, Naïma Kebdani, Chrystèle Charles-Delobel, Yves Bertheau, Angèle Geldreich, Géraldine Coué-Philippe, Annick Diolez, Marie-Noëlle Duplan, Marcel Romaniuk |
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| Přispěvatelé: | Unité de recherche Phytopathologie et Méthodologies de la Détection (PMDV), Institut National de la Recherche Agronomique (INRA), Agrogène SAS, Partenaires INRAE, Institut de biologie moléculaire des plantes (IBMP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA) |
| Rok vydání: | 2005 |
| Předmět: |
DNA
Plant Base pair [SDV]Life Sciences [q-bio] Biology Zea mays 01 natural sciences Genome DNA sequencing Plant Viruses Analytical Chemistry Conserved sequence law.invention QRT-PCR 0404 agricultural biotechnology law Environmental Chemistry Promoter Regions Genetic Polymerase chain reaction DNA Primers Pharmacology Genetics Reverse Transcriptase Polymerase Chain Reaction fungi 010401 analytical chemistry Reproducibility of Results 04 agricultural and veterinary sciences Amplicon Plants Genetically Modified biology.organism_classification 040401 food science Molecular biology 0104 chemical sciences Genetically modified organism DNA Viral Seeds Viruses Cauliflower mosaic virus Oligonucleotide Probes Agronomy and Crop Science Food Science |
| Zdroj: | Scopus-Elsevier Journal of AOAC INTERNATIONAL Journal of AOAC INTERNATIONAL, AOAC International, 2005, 88 (2), pp.547-557. ⟨10.1093/jaoac/88.2.547⟩ |
| ISSN: | 1944-7922 1060-3271 |
| DOI: | 10.1093/jaoac/88.2.547 |
| Popis: | A highly sensitive quantitative real-time assay targeted on the 35S promoter of a commercial genetically modified organism (GMO) was characterized (sF/sR primers) and developed for an ABI Prism® 7700 Sequence Detection System and TaqMan® chemistry. The specificity assessment and performance criteria of sF/sR assay were compared to other P35S-targeted published assays. sF/sR primers amplified a 79 base pair DNA sequence located in a part of P35S that is highly conserved among many caulimovirus strains, i.e., this consensus part of CaMV P35S is likely to be present in many GM events. According to the experimental conditions, the absolute limit of detection for Bt176 corn was estimated between 0.2 and 2 copies of equivalent genome (CEG). The limit of quantification was reached below 0.1% Bt176 content. A Cauliflower Mosaic Virus control (CaMV) qualitative assay targeted on the ORF III of the viral genome was also used as a control (primers 3F/3R) to assess the presence of CaMV in plant-derived products. The specificity of this test was assessed on various CaMV strains, including the Figwort Mosaic Virus (FMV) and solanaceous CaMV strains. Considering the performance of sF/sR quantification test, the highly conserved sequence, and the small size of the amplicon, this assay was tested in a collaborative study in order to be proposed as an international standard. |
| Databáze: | OpenAIRE |
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