Influence of Heat Processing on DNA Degradation and PCR-Based Detection of Wild-Type and Transgenic Maize

التفاصيل البيبلوغرافية
العنوان: Influence of Heat Processing on DNA Degradation and PCR-Based Detection of Wild-Type and Transgenic Maize
المؤلفون: Boris Vishnepolsky, David Mikeladze, Nelly Datukishvili, Kakha Bitskinashvili, Tamara Kutateladze, Inga Gabriadze
المصدر: Journal of Food Quality, Vol 2019 (2019)
بيانات النشر: Hindawi, 2019.
سنة النشر: 2019
مصطلحات موضوعية: Genetically modified maize, Article Subject, Wild type, lcsh:TX341-641, Amplicon, Biology, DNA extraction, Molecular biology, law.invention, Genetically modified organism, law, Genetic marker, Safety, Risk, Reliability and Quality, Gene, lcsh:Nutrition. Foods and food supply, Polymerase chain reaction, Food Science
الوصف: Reliable detection of genetically modified (GM) maize is significant for food authenticity, labelling, quality, and safety assessment. This study aims to evaluate the factors influencing degradation and polymerase chain reaction (PCR) amplification of DNA from the wild type and transgenic maize (events Bt-176 and MON810) during thermal treatment at 100°C and 121°C. A new PCR method was developed targeting the Cry1Ab gene to detect insect-resistant GM plants. The yield of genomic DNAs extracted by the DNeasy plant mini kit dramatically decreased while DNAs obtained by cetyltrimethyl ammonium bromide- (CTAB-) based method did not show any visible changes in the yield by the time of processing. Treatment at 100°C did not significantly affect either genomic DNAs or amplicons. Heating at 121°C induced time-dependent degradation of genomic DNAs and exogenous Cry1Ab gene; however, it did not have any considerable influence on the exogenous 141 bp amplicons or endogenous amplicons in the range of 102 bp to 226 bp with the exception of the event MON810 extracted by the DNeasy plant mini kit. More yield was observed at 226 bp than 140 bp fragment of the invertase gene. The 141 bp fragment of the transgenic CaMV 35S promoter exhibited the highest thermal stability of all the examined amplicons. Analysis of foodstuffs demonstrated 102 bp amplicons specific for the zein gene as the effective marker to detect maize in the processed foods. The obtained results demonstrate that PCR-based detection of the wild type and transgenic maize is dependent on the combination of different parameters of crucial factors such as temperature and duration of exposure, transgenic event, DNA extraction method, DNA marker, and size and location of amplicons.
وصف الملف: text/xhtml
اللغة: English
تدمد: 0146-9428
DOI: 10.1155/2019/5657640
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b73b07255bd2cce011f17a3b9932e13e
حقوق: OPEN
رقم الأكسشن: edsair.doi.dedup.....b73b07255bd2cce011f17a3b9932e13e
قاعدة البيانات: OpenAIRE
الوصف
تدمد:01469428
DOI:10.1155/2019/5657640