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Human infectious diseases and GMO
KOBLIHOVÁ, Tereza
This bachelor thesis is focused on the detection of the transgenes in DNA of the genetically modified plants, especially on the detection of the transgene nptII. Moreover, it will discuss the determinations of the fission ratio (the amount of copies) of the injected transgenes into the next generation. The theoretical part of this bachelor thesis includes information about bacteria because some of their kinds are the reasons of human infection diseases, however, couple of them are used in genetic engineering for plant transformation at the same time. The diseases which are caused by bacteria are cured by antibiotics. Antibiotics are antimicrobic substances which are used to stop the growth of or exterminate these microorganisms. The theoretical part is especially focused on findings related to antibiotics and gene manipulated organisms (GMO). They are related to the concerns about using the selective gene nptII in the construct of these organisms, because this gene codes enzyme neomycin phosphotransferase II, which deactivates the effects of the Antibiotic called kanamycin. Organisms, where this gene is included are getting resistant to these antibiotics. The aim of the methodical part was the cultivation of own genetic modified plants carrying artificially injected genes for the resistance to kanamycin (nptII) and the signal gene GUS. Furthermore, we were looking for the subsequent determination of presence of the gene ntpII in the samples from these plants. The first necessary step to the determination of the gene nptII in the transgenic plants was multiplying of the DNA section containing this gene. That was accomplished by Polymerase Chain Reaction (PCR). After PCR was adapted, the detection of nptII gene in samples was made by gel electrophoresis. A histochemical test was used for the detection of the presence of the gene nptII in the plant samples and the confirmation of the transformation. As a model organism, the Nicotiana tabacum was chosen, which was transformed by Agrobacterium tumefaciens.
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