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Gene expression reporters in \kur{Drosophila melanogaster}.
ŠTROS, Jiří
Drosophila melanogaster is a widely used model organism in genetic research and in a number of other disciplines associated with medical and biotechnological issues. The first part of this thesis presents a review of some basic genetic tools and gene expression reporters used in D. melanogaster research with emphasis on the use of GFP reporter. The second part presents a description and results of my experimental work aimed at the reporter construct consisting of adenosine deaminase gene (ADGF-A) and GFP marker. When using gene expression reporters such as GFP, it is important to know whether the presence of reporter does not affect the studied process. The experiment described in this study tested whether the fusion protein consisting of GFP and adenosine deaminase is fully functional enzyme or whether the enzyme activity is influenced by the presence of fluorescent tag. Results of this work support the usefulness of using the fusion construct as a gene expression reporter.
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Study of Ectonucleotidases and Adenosine Deaminases in Drosophila
PREUER, Kristina
Extracellular adenosine triphosphate and extracellular adenosine are important regulatory molecules in the human immune system. The concentrations of these molecules are in turn regulated by ectonucleotidases and adenosine deaminases. In this thesis I attempt to test the gene silencing efficiency of RNA interference for three different genes coding for such enzymes in the model organism Drosophila melanogaster.
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Effect of oxidative stress on antioxidant enzymes in \kur{Drosophila melanogaster}.
SZAKOSOVÁ, Klára
Oxidative stress reflects an imbalance between reactive oxygen species and antioxidant defense, which can result in cellular damage. The reactive radicals can interact with cell components, including proteins, lipids and nucleic acid. Antioxidant enzymes are the main defense against oxidative stress, representing by two the most important enzymes, catalase and superoxide dismutase. In this thesis I studied the effect of oxidative stress on activity of antioxidant enzymes in Drosophila melanogaster and Locusta migratoria, after exposure to the herbicide paraquat.
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Cold adaptation of insects: the role of shock proteins from Hsp family.
ŠTĚTINA, Tomáš
Heat shock proteins (Hsps) form evolutionarily conserved group of proteins that are best known for their unique function in repair of miss folded proteins that would otherwise aggregate in cells during stress conditions. The most extensively studied is the inducible Hsp70. Using 3rd instar larvae of transgene mutant of fruit fly, Drosophila melanogaster that lacks all copies of hsp70 gene, we assessed: 1) whether the absence of this gene is deleterious for larvae in their capacity to survive cold stress; and 2) whether any other gene from Hsps family may compensate for the loss of hsp70. We found that 1) survival of cold stress is not altered in Hsp- larvae when compared to wild type Hsp+ larvae; 2) there is no other gene of Hsps family that would compensate for the loss of hsp70 at the level of transcription. Our ongoing effort is focusing on analysis of cold stress response at the protein level.
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Long-Term Cold Acclimation Extends Survival Time at 0°C and Modifies the Metabolomic Profiles of the Larvae of the Fruit Fly Drosophila melanogaster
KORBELOVÁ, Jaroslava
We assessed survival of larvae of the fruit fly, Drosophila melanogaster at 0°C after rearing them under three different acclimation regimes. A comparison of the larvae acclimated at constant 25°C with those acclimated at constant 15°C followed by constant 6°C for 2 d showed that long-term cold acclimation extended the lethal time for 50% of the population (Lt50) during exposure to constant 0°C as much as 630-fold (from 0.137 h to 86.658 h). Detailed metabolomic analyses showed that long-term cold acclimation modified the metabolomic profiles of the larvae. The most prominent responses were accumulations of proline (up to 17.7 mM) and trehalose (up to 36.5 mM). In addition, restructuring of the glycerophospholipid composition of biological membranes was observed.
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Study of physiological characteristics of \kur{Drosophila melanogaster} IDGF mutants
KOCIÁNOVÁ, Jitka
Products of Drosophila Idgf genes comprise a small protein family, related to chitinases. The main function of these proteins is to support cell proliferation and activate immune responses. IDGF proteins are present in large quantity in the hemolymph, especially during infections. In this study, I compared the speed of hemolymph cloting, the number and morphological properties of hemocytes after infection by entopathogenic nematodes and parasitic wasps. The results confirm the role of Idgf genes in Drosophila immune system.
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Identification of interacting partners of Discs overgrown in vivo
MAREŠOVÁ, Petra
The mutated forms of the Discs overgrown gene causes overproliferation of imaginal discs of Drosophila melanogaster. Somatic mutations in its human counterpart, casein kinase I epsilon, were strongly associated with human breast cancer. Using the advantage of a high conservancy between fly's dco and human casein kinase I epsilon genes we have chosen D. melanogaster as a model organism to provide a list of probable Dco interaction partners via tandem affinity purification and mass spectrometry analysis. However, these proteins need to be independently verified as true Dco interaction partners.
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