HSBMB Newsltr - GMO-detection - Hellenic Society of...

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Hellenic Society of Biochemistry and Molecular Biology Newsletter, (1999) 46: 48-56. Cloned-DNA Detection in Raw and Processed Food and Feed Derived from Genetically Modified Plants Alexios N. Polidoros and Athanasios S. Tsaftaris Dept. of Genetics and Plant Breeding, Aristotelian University of Thessaloniki email A.N.P: alexios@agro.auth.gr, A.S.T: tsaft@agro.auth.gr Abstract With the increasing availability of genetically modified plant products, it has become necessary to develop easy routine techniques for identification of such products in order to follow labeling requirements and other EU and National regulations. Widely applicable PCR-based screening methods might be most adequate and effective. Sequences commonly found in many transgenic crops, such as the CaMV-35S promoter, the nptII antibiotic resistance gene, or the nos - terminator may provide a primary target for screening. Alternatively, PCR-based identification may be directed to the modification-specific gene. In this study, we developed specific primer pairs and tested commercial kits for detection of cloned DNA in fresh and processed material. Introduction Advances in plant biotechnology and their application in plant breeding permitted the creation of new transgenic plant varieties and their wide use in modern agriculture. Introduction of foreign genes into plant genomes presents potential safety risks for man and the environment, and commercial use of Genetically Modified Organisms (GMO) and their products is regulated by relevant legislation in European Union and consequently in our country. Deliberate release of GMOs to the environment is regulated according to EU directive 220/90 and labeling of raw material, food and feed derived from GMOs, is enforced by directives 1813/97 and 1139/98. This creates the need for development of easy routine methods for detection of GMOs in raw material, and processed food and feed. Detection of the modification in Genetically Modified Plants (GMPs) can be targeted to the DNA construct integrated into the host genome, the products of the incorporated genes at the mRNA and protein level, or the identification of the novel phenotype conferred by the modification. Although in fresh plant material detection can be efficient for all the above targets, in processed material the only available target is (in most of the cases) the inserted DNA. DNA-sequences introduced into the plants by genetic engineering can be amplified and detected using the polymerase chain reaction (PCR) (Saiki et al., 1985). Target sequences available for screening may be common in many transformed varieties. Such sequences are present in genetic elements most commonly found in transgenic crops, including the neomycine-3'-phosphotransferase ( npt II) marker gene, the cauliflower mosaic virus 35S promoter (CaMV-35S) or the nos-terminator ( nos -3'). The modification-specific gene, which is unique for each transgenic variety could be conclusively identified through a gene-specific
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This note was uploaded on 09/13/2010 for the course DGPB 024e taught by Professor Alexiospolidoros during the Spring '10 term at Aristotle University of Thessaloniki.

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HSBMB Newsltr - GMO-detection - Hellenic Society of...

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