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Application for the Data Processing in the Area of Genome Engineering
Brychta, Jan ; Burgetová, Ivana (referee) ; Očenášek, Pavel (advisor)
This masters thesis has a few objectives. One of them is to acquaint with the problems of genome engineering, especially with fragmentation of DNA, the macromolecule DNA, the methods for purification and separation of the nucleic acids, the enzymes used for modification of these acids, amplification and get to know with cluster and gradient analysis as well. The next aim is to peruse the existed application and compare it to the layout of the proposed application, that is the third aim. The last one from the objectives is the implementation and the report how was the application tested by the real data. The results will be discussed as well as the possibilities of the further extension.
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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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Gene engineering methods in Leishmania research
Kriegová, Gabriela ; Bečvář, Tomáš (advisor) ; Arbonová, Lenka (referee)
Gene engineering methods are now increasingly used in the study of leishmania. These modern DNA technologies make it possible to manipulate with the genome of organisms, allowing it to be altered and repaired. The most important methods include CRISPR/Cas9, RNA interference (RNAi), dimerizable Cre recombinase (DiCre), plasmid shuffle and gene tagging. The choice of method depends on whether essential or non-essential genes are being studied as well as the reason of the study. These methods investigate leishmania using a reverse genetics approach that allows the phenotype to be studied. This work summarizes the current knowledge on the use of gene engineering methods in Leishmania, the necessary modifications of the procedures and discusses their limitations and advantages.
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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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Geneticky modifikované plodiny a možnosti jejich využití - případová studie v České republice
Pillař, Miroslav
This thesis deals with the topic of genetically modified crops and possibilities of their use in the Czech Republic. The attitude of randomly selected group of Czech citizens toward this technology was evaluated by using the method of semi-standardized interviews and questionnaires. The aim of this work was to document actual situation related to GMO and other usage of modified crops in the Czech Republic. From the results it can be seen that the interest of agricultural subjects regarding mentioned technology is decreasing rapidly. The awareness of respondents was mostly of negative character due to confusion about its effects on human health and the environment. On the contrary the scientific community sees the main problem in environmental and biodiversity threats. Due to the lack of long-term scientific researches it is currently impossible to confirm or disprove advantages or disadvantages brought by this technology. Thus further conclusive scientific studies are needed.
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Knowledge and Opinions of Students on Genetically Modified Organisms
Semencová, Barbora ; Hlaváčová, Lucie (advisor) ; Vojíř, Karel (referee)
This diploma thesis is focused on topic of genetically modified organisms and their use in the practical sectors of human life. Theoretical part of the thesis defines general terms GMO, plasmid, genetic engineering, biotechnology. It also records historical milestones relating to the problematic, deals with individual techniques of genetic engineering and briefly states legislative procedures in context of dealing with GMO. It gives examples of transgenic organisms and summarizes advantages and disadvantages of their use.Practical part of the thesis contains educational program called "Genetically modified organisms", which was conceived by the author and includes a draft of a lesson inclusive of teaching materials - powerpoint presentations, worksheets, interactive worksheets, auxiliary text for teacher and written preparation. Research part deals with high school students change of view about using GMOs after completing the educational program. Due to analysis was proven that most of the attitudes and knowledge about GMO was changed after completing the educational program (for example in issues of willingness to consume GM food and animal products, perception of advantages and disadvantages etc.) Data was still unchanged in questions which cannot be affected by the program (control of food packaging or...
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PCR detection of genetically modified soybean in food and feed
CHADIMOVÁ, Nikola
Genetically modified organisms which genetic equipment has been specifically targeted of have been acquiring properties they did not have before. One of the main reasons for this aimed genetic modification in plants is herbicide resistance. Especially in soy case is used the modification to resist to glyphosate. The safety of the food which has been made of genetically modified crops or products of animal fed with GM feed has not been proved so far. This bachelor thesis focuses on methods for detection of GMOs in food and feed. Also, it focuses on verifying the data concerning the presence of GMOs presented by companies on food or feed packaging. The research part of this work contains information about genetic engineering, the issue of genetically modified organisms and also focuses on the description of techniques needed to detect GMOs, especially in processed foods, but also in feeds available on the Czech market. The methodical part of this bachelor thesis focuses on the detection of CaMV 35S promoter occurring in DNA crops that have undergone genetic modification. The presence of a specific transgenic material involved the detection of a particular glyphosate-resistant gene, which is particularly known under the trade name Roundup. There were used basic molecular biological methods to analyze GMOs.
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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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