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Science Fiction and the Prediction of Technology, Past and Present
Skorobogatova, Margarita ; Sedláček, Pavel (referee) ; Froehling, Kenneth (advisor)
Cílem této bakalářské práce je prozkoumat, zda-li sci-fi dokáže predikovat budoucí vývoj technologií. První kapitola představuje sci-fi spisovatele 19. a 20. století, kteří předpovídali řadu technologických vynálezů vyskytujících se v současnosti. Druhá kapitola se zabývá sci-fi kinematografií, která je popsána a doložena příklady z sci-fi filmů a televizních seriálů. Poslední kapitola je základem různorodých technologických vynálezů současnosti, které jsou popsány a demonstrovány režiséry sci-fi filmů, a považovány za nejvíce pravděpodobné přiklady, které můžou být realizovány v blízké budoucnosti. Závěr popisuje výsledky provedených výzkumů a definuje případný vývoj v budoucnu.
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Birth
Piskova, Olena ; Klímová, Barbora (referee) ; Gabriel, Michal (advisor)
The thesis is a continuation of my subject, "Why?", Also includes the theme of man and the environment. I started to be interested in the controversial topic of genetically modified organisms, genetic engineering. I found a personal story in it. Now it is difficult to comment on the extent to which these researches are important, but we shouldn’t stop there.
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Production of human milk oligosaccharides in the cell factory of E. coli
Havrdová, Jana ; Bojarová, Pavla (advisor) ; Smrček, Stanislav (referee)
Human milk oligosaccharides (HMOs) are among the most abundant components of human breast milk and are essential for the development of neonatal health. It is very challenging to isolate these oligosaccharides from animal milk, especially the less abundant ones. Therefore, different approaches are to be sought after. Chemical and enzymatic syntheses of these compounds are labor-intensive, and expensive affording low yields. A newly adopted approach to HMO synthesis is through bac- terial cell factories, in which genetically engineered bacterial strains can use cheap carbohydrate substrates and convert them into specific oligosaccharides. The aim of this thesis is to examine the feasibility of using selected bacterial strain (E. coli) for the production of HMOs. With lactose as a glycan substrate, the bacterial host has to be β-galactosidase deficient, otherwise, the substrate would be degraded. In order to generate higher lactose intake in the cell, a crp gene that encodes for the positive transcriptional regulator (Catabolite Activator Protein - CAP) can be incorporated into the host organism. The cloned plasmid - pRSFDuet- 1-crp was used for the transformation of into the selected bacterial strain. Lactose from the cells was purified by gel chromatography and the influence of CAP over expression...
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Preparation and characterisation of mRNA/DNA transfection vectors
Horák, Tomáš ; Chmelíková, Larisa (referee) ; Skopalík, Josef (advisor)
This diploma thesis deals with genetic engineering, especially the transfection of DNA into MSCs (Mesenchymal stromal cells) and dendritic cells. Both lipoplexes and metal magnetic nanoparticles were tested to introduce the vector into cells. The research was focused on finding more efficient methods of transfection. According to analysis on MADLS and gel electrophoresis, aspects playing an important role in conjugation and subsequent transfection were found. Conjugation occurs after only 4 minutes, as evidenced by an increase in zeta potential, but to achieve full conjugation it is necessary to incubate the sample for 20 minutes. Incomplete conjugation to iron nanoparticles resulted in strong carrier-carrier interactions, which formed an unwanted conglomerates. Encapsulation into liposomes with cationic surface treatment was without complications. The success rate of GFP-labeled protein expression after transfection by these methods was calculated to be 95%, resp. 91%. This result is due to low cytotoxicity. However, commercial tested kits on dendritic cells had a success rate below 5% with high cytotoxicity.
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Preparation and characterisation of mRNA/DNA transfection vectors
Horák, Tomáš ; Chmelíková, Larisa (referee) ; Skopalík, Josef (advisor)
This diploma thesis deals with genetic engineering, especially the transfection of DNA into MSCs (Mesenchymal stromal cells) and dendritic cells. Both lipoplexes and metal magnetic nanoparticles were tested to introduce the vector into cells. The research was focused on finding more efficient methods of transfection. According to analysis on MADLS and gel electrophoresis, aspects playing an important role in conjugation and subsequent transfection were found. Conjugation occurs after only 4 minutes, as evidenced by an increase in zeta potential, but to achieve full conjugation it is necessary to incubate the sample for 20 minutes. Incomplete conjugation to iron nanoparticles resulted in strong carrier-carrier interactions, which formed an unwanted conglomerates. Encapsulation into liposomes with cationic surface treatment was without complications. The success rate of GFP-labeled protein expression after transfection by these methods was calculated to be 95%, resp. 91%. This result is due to low cytotoxicity. However, commercial tested kits on dendritic cells had a success rate below 5% with high cytotoxicity.
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Evolutionary and genetic engineering of bacterial producers of polyhydroxyalkanoates
Nováčková, Ivana ; Patáková, Petra (referee) ; Koutný, Marek (referee) ; Obruča, Stanislav (advisor)
This doctoral thesis deals with the topic of evolutionary and genetic engineering of polyhydroxyalkanoates (PHA) producing bacteria. Apart from these topics, the issue of biotechnological production of PHA on model hydrolysates of lignocellulosic biomass with the use of extremophilic microorganisms is also studied, as well as the development of an alternative method of PHA isolation. The themes were freely linked to previous experiments and reflected the currently solved projects in a working group. Doctoral thesis is prepared in the form of a commented discussion of published works, which are part of it in the form of appendices. Evolutionary engineering was mainly applied to the model PHA producing bacterial strain Cupriavidus necator H16. By adaptation to levulinic acid, isolates producing copolymer P(3HB-co-3HV) with a higher content of the 3HV fraction were obtained, which leads to improved properties of the polymer for further processing. As well as culture growth also the amount of total PHA in the biomass was higher. By long-term adaptation of the same strain to osmotic stress and the presence of copper ions, the isolates which are characterized in the second publication, were obtained. Based on obtained data, it was possible to observe differences in the adaptation process, where the adaptation to osmotic stress was gradual, while a significant step in the increase of biomass and PHA signaling faster adaptation was observed for copper. Based on the analyses, the significant role of PHA in the adaptation of the C. necator H16 strain to the tested stressors was discussed, it did not consist only in the increase in the amount of polymer in the biomass, but also in enhancement of whole PHA cycle, which also leads to an increase of the pool of monomeric units showing protective functions. By adaptation to -captolactone, a unique precursor of 4HB, the copolymer P(3HB-co-4HB) was obtained. The properties of this copolymer are again more favorable than of the homopolymer P(3HB), even with a low content of 4HB, which we also achieved in a laboratory bioreactor. A further increase in the 4HB fraction could be achieved using deletion mutants with the absence of relevant genes, which is discussed more in the text. The production of PHA on models of lignocellulosic biomass hydrolysates originating from, for example, the food industry was tested in combination with the use of extremophile producers, when the preference of the contained monosaccharides (hexoses, pentoses) for individual producers was discussed. For the purpose to get closer to real hydrolysates, the resistance of the strains to relevant potential microbial inhibitors was also tested. The susceptibility of halophilic and thermophilic PHA producers to osmotic stress was used in the development of an alternative isolation approach that would reduce the economic and ecological burden of the process compared to standard extraction using chlorinated solvents. Application of SDS detergent at low concentrations while simultaneously exposing the cells to higher temperatures led to the gain of high purity polymer without loss of yield. The recycling process of used SDS is also a possibility.
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PCR Detection of GMO in Food and Feed
ŠKRNOVÁ, Dominika
The term "genetically modified organism (GMO)" has become a controversial topic in many countries as its benefits for both food producers and consumers are accompanied by potential biomedical risks and environmental side effects. In this thesis, I attempt to summarize up-to-date knowledge about genetically modified (GM) crops. I also introduce different methods for detecting GMOs in food and feed. In the practical part of this thesis, thirty food and feed samples containing maize were randomly collected from different sources. The aim of this study was to test these samples for the presence of GMO, verify the information given on the label, and check that it complies with the EU legislation requiring that any food or feed containing more than 0.9% GM content has to be labeled. The next aim was to optimize the method for DNA extraction from the tested samples. In order to isolate genomic DNA from the samples, I used the NucleoSpin Food kit and the MagCore Automated Nucleic Acid Extractor. The concentration of extracted DNA was evaluated by ultraviolet (UV) absorption using a BioSpec-nano spectrophotometer. The gene coding for the zein storage protein of maize was used to confirm the presence of amplifiable maize DNA. DNA extraction was followed by PCR protocols using different sets of primers to detect four maize events: Bt11, Bt176, Mon810, and T25. Agarose gel electrophoresis was used for the separation of DNA fragments that were subsequently visualized using InGenius manual gel documentation system. In spite of the curent regulations, none of the three samples tested positive for the presence of MON810 maize had any such indication on their label. The samples that claimed to be "GMO free" on their label actually did not show the presence of any of the GM maize events tested.
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Potential of genetic engineering for breeding plants tolerant to abiotic stresses: cold resistance in rice
Lotová, Gabriela ; Lipavská, Helena (advisor) ; Janská, Anna (referee)
With increasing population and climate change, there has been an increase in efforts to breed more efficient crops. Genetic engineering has opened unprecedented breeding possibilities in developing plants with desired traits. Transgenic crops with better qualities, including resistance to adverse environmental conditions, can contribute to solving problems of hunger and malnutrition in developing countries. Although society perceives genetically modified crops rather negatively, these crops are widely used as feed for livestock and outside Europe also for human nutrition. Because of the complexity of resistance to abiotic stress, the utility of genetic manipulations for the breeding of resistant plants was previously not anticipated. However, it turned out that modification of the stress signalling cascade or transcription factors can lead to success. This thesis summarizes the possibilities of genetic modification of crops, which may result in better tolerance to cold, and is mainly focused on rice. Part of the work deals with transduction of cold signal, whose modification can also result in increased tolerance to cold. Another part deals with transcription factors that activate expression of low temperature- resistant genes. The greatest attention is paid to CBF/DREB transcription factors that...
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Genetically modified plants in practical applications
Říhová, Barbora ; Fischer, Lukáš (advisor) ; Cvrčková, Fatima (referee)
Genetic engineering (GI) of plants is a very current topic, and more and more controversial, since it is becoming an inseparable part of our lives. GI has, among other things, a great potential to help solve the current problem of hunger and malnutrition in certain parts of the world. The goal of this project is to clarify what genetically modified (GM) plants are, to present the possibilities of their practical use, to explain methods of preparation and to consider their advantages and eventual risks. By GM plant we understand a plant whose genetic information has been changed by introducing or removing part of the genetic information (a section of DNA, generally a gene). The most frequently used methods of transformation are the transformation by Agrobacterium tumefaciens and the balistic method. GM plants can be used for production of food or feed in better quality and greater quantity, thanks to an introduction of higher resistance, whether it is to biotic stress (pest, virus or bacterial resistance) or to abiotic stress (drought, salinity, toxic substance in the ground..). In particular, the resistance to abiotic stress is an important issue these days, since through the climate changes some regions suffer more and more from insufficient precipitation and consequent drought. Some of the GM plants grown...
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Genome editing using programmable endonucleases
Hanečková, Radmila ; Sedláček, Radislav (advisor) ; Sýkora, Michal (referee)
Programmable endonucleases are engineered proteins that recognize specific nucleotide sequences and that are capable of introducing double-strand breaks within these sequences. Zinc-finger nucleases have been used extensively as a tool in genome editing, the practice of introducing changes into genomes of cell lines or whole organisms as a way to study gene function. Recently, new types of programmable endonucleases have emerged in the form of transcription activator like effector (TALE) nucleases and the CRISPR/Cas system. The types differ in respect to their mechanism of function, accessibility, selectivity, frequency of off-target cleavage and cytotoxic effects. Here, we compare zinc-finger nucleases, TALENs and the CRISPR/Cas system and explore their current and possible future applications in a broad spectrum of research ranging from developing genetically modified organisms to gene therapy. Powered by TCPDF (www.tcpdf.org)
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