Detection of genetically modified insect resistant tomato via polymerase chain reaction

Download
2004
Sönmezalp, C. Zeynep
Tomato, which is one of the most important component of human diet, has been genetically modified to develop some properties like delayed ripening and insect resistance. In order to give a choice to the consumer, it is necessary to detect and label GM foods. This study was carried out to detect genetically modified tomato samples purchased from different food markets of Turkey. PCR method was used to detect genetically modified insect resistant tomatoes. The DNAs of collected samples were isolated according to CTAB DNA extraction protocol and the amplification capacity of isolated samples were checked with patatin specific control PCR. Screening tests of tomatoes were done by targeting 35S promoter, Nos terminator and NptII kanamycin resistance gene with four primer sets. It was aimed to detect Cry1A and Cry1Ac genes with three PCR systems, in order to identify insect resistant samples. From twenty-eight samples, twenty-two gave positive amplification signal in NptII specific PCR system and results were confirmed with sequence analysis. Additionally, we observed seventeen and ten DNA fragments with Cry1A-F/Cry1A-R and Cry1Ac-F/Cry1Ac-R primer sets respectively, it is necessary to confirm these results with DNA sequencing.

Suggestions

Detection of genetically modified maize via polymerase chain reaction
Aydın, Gamze; Gürakan, Candan; Department of Biotechnology (2004)
In recent years, foods produced by genetic engineering technology have been on the world food market. The biosafety aspects, regulations, and labelling of these foods are still contentious issues in most countries. It is necessary to have approval for the use of GMOs in the production of food. Thus, detection and quantification of GMOs play crucial role for developing regulations on GM foods. In this study, raw and processed maize samples were analysed for genetic modification using a DNA based detection me...
Screening for genetically modified tomatoes & tomato seeds and identification of CRY1AC and SAM-K specific modifications using gene and construct specific PCR
Uçkun, Esra; Gültekin, Güzin Candan; Department of Biotechnology (2007)
This study was carried out to analyze tomato samples and tomato seeds, purchased from different food markets of Turkey randomly, for the presence of genetic modification by using PCR method as it allows more specific detection. The DNAs of collected samples were isolated according to CTAB DNA extraction protocol and also with extraction kits. Screening tests of tomatoes were done by targeting 35S promoter, NOS terminator and NptII kanamycin resistance gene with eight different primer sets. Real time PCR is ...
Analysis of self-processing mechanism of galactose oxidase by site-directed mutagenesis and heterologous expression in Escherichia Coli
Gençer, Burçak; Ögel, Zümrüt Begüm; Department of Biotechnology (2005)
In this study, self-catalytic maturation of heterologously expressed pro-galactose oxidase was analysed in E.coli by altering some amino acids which were supposed to play a crucial role in pro-peptide removal. Galactose oxidase (GOase; EC 1.1.3.9) from Fusarium graminearum; having a molecular mass of 68kDa, is a monomeric, copper containing enzyme with an unusual thioether bond. The enzyme is produced as a precursor with an additional 8 amino acid pre- and a 17- amino acid pro-sequence at the N terminus. Pr...
Identification and analysis of genomic regions with large between-population differentiation in humans
Myles, S.; Tang, K.; Somel, Mehmet; Green, R. E.; Kelso, J.; Stoneking, M. (Wiley, 2008-01-01)
The primary aim of genetic association and linkage studies is to identify genetic variants that contribute to phenotypic variation within human populations. Since the overwhelming majority of human genetic variation is found within populations, these methods are expected to be effective and can likely be extrapolated from one human population to another. However, they may lack power in detecting the genetic variants that contribute to phenotypes that differ greatly between human populations. Phenotypes that...
Transcriptional analysis of hydrogenase genes in rhodobacter sphaeroides O.U.001
Doğrusöz, Nihal; Gündüz, Ufuk; Department of Biology (2004)
In photosynthetic non-sulphur bacteria, hydrogen production is catalyzed by nitrogenases and hydrogenases. Hydrogenases are metalloenzymes that are basically classified into: the Fe hydrogenases, the Ni-Fe hydrogenases and metal-free hydrogenases. Two distinct Ni-Fe hydrogenases are described as uptake hydrogenases and bidirectional hydrogenases. The uptake hydrogenases are membrane bound dimeric enzymes consisting of small (hupS) and large (hupL) subunits, and are involved in uptake and the recycling of hy...
Citation Formats
C. Z. Sönmezalp, “Detection of genetically modified insect resistant tomato via polymerase chain reaction,” M.S. - Master of Science, Middle East Technical University, 2004.