Development of analysis methods for cry1ac and sam-k gene lines in tomato using pcr and real-time pcr

Download
2010
Uygun, Sahra
Genetically modified organisms are entering the human diet in all over the world. In order to have transparency in the foods that are being consumed, there is a need to trace the genetically modified organisms (GMOs) in the market and consequently this need brings the necessity of analytical methods that are capable of detecting, identifying and quantifying the transgenic events. These analytical methods also form the basis of the labeling regulations that are tried to be formed regarding GMOs. The main aim of this study is to develop and apply the detection methods for the two of the tomato events, delayed ripening and insect resistant. Currently the only validated detection methods are mainly for the corn, soybean, and cotton. There is no validated detection method for tomato. Tomato is one of the most consumed food products in Turkey and it is also among the controversial organisms in terms of genetic modifications and labeling, therefore the analysis of the genetic modifications in tomato is crucial. In this study, DNA-based detection is performed, with PCR being the chosen method of study. In order to detect the GMO-derived DNA, the method of analysis includes the following studies: species-specific, screening, gene-specific, construct-specific and inverse PCR. In addition, the quantification method is developed using the real-time PCR. In order to develop the procedure of identification method, the reference samples are used and the unknown varieties that are to be analyzed using this method are expected to have similarities with the authorized transgenic events.

Suggestions

Development of a loop mediated isothermal amplification (LAMP) based detection platform for genetically modified organism (GMO) detection
Moğol, Ayça Nazlı; Öktem, Hüseyin Avni; Department of Biotechnology (2019)
Genetically modified organisms (GMOs) are being widely used worldwide. Every country has a different legislation regarding the allowed events and GMO levels. This creates the great need for GMO detection. In this study, LAMP assay was used for GMO detection owing to its high sensitivity with the genetically modified organisms; Bt11 maize, GT73 Roundup Ready canola, and transgenic Nicotiana tabacum, targeting the sequences most commonly used in GMO constructions; 35S promoter and Figwort mosaic virus (FMV) s...
A sandwich-type DNA array platform for detection of GM targets in multiplex assay
Cansiz, Sena; Özen, Can; Bayrac, Ceren; Bayrac, A. Tahir; Gül, Fatma; Kavruk, Murat; Yilmaz, Remziye; EYİDOĞAN, FÜSUN; Öktem, Hüseyin Avni (2012-09-01)
Over the last decade, array detection has been developed to qualitatively assess the presence of genetically modified organisms (GMOs). To date, DNA array systems have the highest capabilities as a result of GMOs analysis. We describe the construction of an array platform in the sandwich hybridization format for the detection of transgenic promoter of Cauliflower mosaic virus (CaMV; p35S). Sequence-specific signal development has been achieved by a sandwich complex composed of a surface immobilized capture ...
Development of rapid dipstick assay for food pathogens, Salmonella, by optimized parameters
Çam, Dilek; Öktem, Hüseyin Avni (Springer Science and Business Media LLC, 2018-10-25)
Salmonella is among the very important pathogens threating the human and animal health. Rapid and easy detection of these pathogens is crucial. In this context, antibody (Ab) based lateral flow assays (LFAs) which are simple immunochromatographic point of care test kits were developed by gold nanoparticles (GNPs) as labelling agent for Salmonella detection. For that purpose some critical parameters such as reagent concentrations on the capture zones, conjugate concentrations and ideal membrane type needed f...
Effects of carbon sources and feeding strategies on human growth hormone production by metabolically engineered pichia pastoris
Açık, Eda; Çalık, Pınar; Department of Chemical Engineering (2009)
In this study, effects of different carbon sources and their feeding strategies on recombinant human growth hormone (rhGH) production by Pichia pastoris were investigated by means of cell growth, recombinant protein production and expression levels of hGH and alcohol oxidase (AOX) genes. In this content, firstly, the strain to be used for high level rhGH production was selected between the two phenotypes, i.e., P. pastoris hGH-Mut+ and P. pastoris hGH-MutS. In this selection both phenotypes were compared in...
Development and validation of a resistance and virulence gene microarray targeting Escherichia coli and Salmonella enterica
Davis, Margaret A.; Lim, Ji Youn; Soyer, Yeşim; Harbottle, Heather; Chang, Yung-Fu; New, Daniel; Orfe, Lisa H.; Besser, Thomas E.; Call, Douglas R. (Elsevier BV, 2010-07-01)
A microarray was developed to simultaneously screen Escherichia coli and Salmonella enterica for multiple genetic traits. The final array included 203 60-mer oligonucleotide probes, including 117 for resistance genes, 16 for virulence genes, 25 for replicon markers, and 45 other markers. Validity of the array was tested by assessing inter-laboratory agreement among four collaborating groups using a blinded study design. Internal validation indicated that the assay was reliable (area under the receiver-opera...
Citation Formats
S. Uygun, “Development of analysis methods for cry1ac and sam-k gene lines in tomato using pcr and real-time pcr,” M.S. - Master of Science, Middle East Technical University, 2010.