National Repository of Grey Literature 3 records found  Search took 0.00 seconds. 
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.
Characteristics and gene expression levels of plant transformants modified by one related constructs shearing a gene for proteinase inhibitor.
BEZUNK, František
Tobacco plants were transformed by bacteria Agrobacterium tumefaciens bearing a plasmid construct with a fusion gene SPI2:GFP and selectable marker NPTII coding for resistance of transformants to antibiotic kanamycin. SPI2:GFP gene was created by a fusion of SPI2- (for serine protease inhibitor) and GFP gene (coding for a green fluorescent protein - GFP) sequences. Transformed tobacco plants were selected on a medium with kanamycin. Positively selected plants were selfed and segregation ratios for NPTII determined for each of genotypes. The presence and function of transgenic DNA was verified by methods of PCR, RT-PCR, Western blot and using the stereomicroscopic fluorescence study of transformants of T0 and T1 generations. This Transgenic genotypes of tobacco Nicotiana tabacum, cv. Petit Havana, SR1 WT showing the expression of the fused gene SPI2:GFP on a level of mRNA and GFP protein were obtained. Thesis was perfomed with a support of the project No. M106030 of Ministry of Education, Youth and Sports of the Czech Republic.
The molecular analysis of transgenic \kur{Lycopersicon esculentum} plants harbouring incorporated pmi gene for phosphomannose isomerase
PŘIKRYLOVÁ, Pavla
The aim of this study was to analyze number of incorporated T-DNA harbouring pmi and nptII transgenes and expression of the pmi gene in tomato transformants using Southern blot and Northern blot methods. The presence of a functional PMI protein was assesed using PMI-assay with Chlorophenol red dye. Mannose selection of tomato seeds was established and segregation patterns in T1 progeny were studied.

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