National Repository of Grey Literature 44 records found  beginprevious15 - 24nextend  jump to record: Search took 0.01 seconds. 
Oxidative stress in bacteria - with an emphasis on the model organism of Escherichia coli
Moravcová, Andrea ; Lichá, Irena (advisor) ; Čáp, Michal (referee)
Most bacterial species encounter aerobic conditions during their life, which can be toxic. This leads to a state of oxidative stress. Toxicity of aerobic environment results from the chemical properties of molecular oxygen and its reactive species (ROS). Bacteria had to adapt to these conditions in the past to ensure preservation and prosperity. This thesis is preferably focused on oxidative stress adaptations in the most elaborated bacterial model - Escherichia coli. Regulation of adaptations at the regulation of transcription, translation and metabolism level are described with emphasis on molecular mechanisms. The main adaptation mechanism against oxidative stress is the deactivation of ROS, as well as the repair of damaged cell structures (macromolecules). These enzyme activities are regulated by several transcriptional regulators. The transcriptional regulators OxyR and SoxRS have been well studied in E. coli. Even though these regulators are conserved across the bacterial spectrum, they may not have the same function in all organisms. For this reason, also other, more or less studied bacterial species - Bacillus subtilis, Streptomyces coelicolor, Pseudomonas putida, Pseudomonas aeruginosa - were included in this thesis. The various strategies of how these bacteria use not only OxyR and SoxRS...
Preparation and validation of a system for the study of regulation of gene expression of yeast linear cytoplasmic plasmids
Horáčková, Kamila ; Vopálenský, Václav (advisor) ; Čáp, Michal (referee)
There is currently very few information about the transaltion of linear cytoplasmatic plasmids occured in yeast cells Kluyveromyces lactis. However, there is a relatively well developer information about their transcription apparatus. A study of transkript linear plasmids revealed an atypical organization at the 5ʼ end. Those ends contain nontemplate polyadenylation and they are missing the N7 methylguanosine hat. Because of the presence of this structure, which is localized at 5ʼend of plasmids specific mRNA, raised a question regarding the iniciation of the translation. The present thesis is focused on the preparation of reporter systém suitable for studying the influence of a number of the nontemplate adenosins, which were added at the 5ʼ ends of mRNA linear plasmids. The frist step was making a construction of dual yeast cell plasmids carring two reporters genes, which are under the controle of two different promoters. After a successfull construction, the aktivity of promoters TEF1 and PGK1 was measured, whereby the promoter TEF1 proved twice stronger. The transcription start site of both promotor was determined. The second step was the construction of a reporter system directly in yeast cell plasmid pGKL. Reporter genes were under the controle of two promoters originating from the pGKL...
Differentiation in Saccharomyces cerevisiae colonies
Čáp, Michal
Yeast colonies are, alongside to bacterial biofilms, multicellular communities formed by unicellular microorganisms. These specific communities differ in many ways from populations cultivated in planktonic cultivations. Gradients of nutrients, metabolic by- products and other factors are formed and preserved within these structures, which provides a basis for cellular differentiation. Current literature concerning these issues with emphasis on yeast colonies and biofilms is summarised in the Introduction of this work. Section Results then describes my contribution to the knowledge on the differentiation of the colonies of Saccharomyces cerevisiae as a model system for studying microbial multicellular structures. Previously described horizontal differentiation, i.e. differentiation between colony centre and margin, is further characterised with respect to ammonia signalling and stress resistance. The importance of genes conferring the cell`s oxidative stress resistance in colony differentiation was studied and it was concluded that not stress resistance but rather metabolic and other changes promoted by ammonia signal are important for colony differentiation and survival. A new type of colony differentiation - the horizontal, i.e. differentiation between upper and lower part of the colony, is...
Codon usage and isoacceptor tRNA expression as a mechanism to control gene expression
Daumová, Lenka ; Čáp, Michal (advisor) ; Štěpánek, Luděk (referee)
Genetic code is defined as a set of rules, which encode aminoacid sequences in proteins, according to codon usage. It is widely known, that there are multiple codons for most aminoacids, thanks to the degeneracy of the genetic code. There was a hypothesis, that silent mutations, which result in a synonymous codon and not in incorporating of a different aminoacid into the peptide chain, don't affect the gene expression. Later however, it was found through more detailed research on molecular level, that codon usage bias is in fact one of the factors, that regulate translation effectivity and rate, mRNA stability or even protein folding and gene expression. There has been many studies published on these topics. This bachelor thesis is a review of these studies. First, I summarize basic information on tRNA, its structure and modifications in anticodon loop. Next I write about base pairing between codon and anticodon, including the non-canonical wobble base pairing. Then I emphasize on codon bias, its causes, its relationship with genome GC content. I also include some author's conjectures about how to approach this phenomenon, which codons are optimal and what is the impact of codon usage bias on translation efficiency. I cite many studies on this topis, which was researched on many model organisms,...
Military history in interwar Czechoslovakia
Cáp, Michal ; Šedivý, Ivan (advisor) ; Beneš, Zdeněk (referee)
1 Annotation This work tries to tackle concept of history of knowledge and possibilities of its application for study of history of military historiography. With the use of this concept, the work will analyse processes of organisation, creations and spread of military historiography in interwar Czechoslovakia. It tries to come to terms with historiographical definition of military history but also its contextualisation considering history of knowledge and sociocultural factors describing the relationship of warfare and society. The work attempts it on the example of main, institutional centres of knowledge management of military history in first Czechoslovak republic. 2
Role of yeast WSS1 protease in DNA repair.
Adámek, Michael ; Grantz Šašková, Klára (advisor) ; Čáp, Michal (referee)
Sustaining the integrity of DNA throughout the lifetime is critical for every living organism. Therefore organisms evolved numerous ways to detect and repair different types of DNA damage caused by various endogenous and exogenous factors resulting in replication stress. Defects in these repair mechanisms can lead to severe human diseases such as neurological disorders, familial cancers or developmental syndromes. In presented master thesis, we investigated the function of a yeast protein named Wss1, a metalloprotease that participates in a recently discovered DNA repair pathway that proteolytically removes DNA-protein crosslinks. Wss1 shows strong negative interaction with another DNA repair protease, Ddi1, in which case was discovered, that double-deleted yeast strain lacking WSS1 and DDI1 is hypersensitive to hydroxyurea. Hydroxurea is a ribonucleotide reductase inhibitor that, in the end, arrests cells in the S-phase of cell-cycle. Based on previous studies, we performed rescue experiments with various deletions and single-site mutants of Wss1p to assess the involvement of particular yeast Wss1p domains in the replication stress response to hudroxyurea.
The relationship between splicing and posttranslational modifications of chromatin in Saccharomyces cerevisiae
Kovaľová, Libuša ; Folk, Petr (advisor) ; Čáp, Michal (referee)
Protein Prp45, the yeast ortholog of the human transcription coregulator SNW1/SKIP, has been previously associated only with the regulation of pre-mRNA splicing. However, our laboratory found that protein Prp45 genetically interacts not only with the proteins involved in pre-mRNA splicing, but also with factors important for transcription elongation and with chromatin modifying enzymes. Our data and the information about the human ortholog SNW1/SKIP suggest that Prp45 could serve as a regulator coupling splicing, transcription and chromatin state in S. cerevisiae. The main aim of this diploma thesis was to find out whether the protein Prp45, which is essential for cotranscriptional assembly of the spliceosome, affects posttranslational modifications of chromatin on transcribed genes. Using chromatin immunoprecipitation, the influence of prp45(1-169) mutation on trimethylation of histone H3 at lysine 4 and acetylation of histone H3 at lysines 9, 14 and 18 on transcriptionally active genes was not confirmed. The other aim was to analyse the behavior of cells synchronized by α-factor by using flow cytometry. According to our results, prp45(1-169) mutation leads to the prolongation of the cell cycle. For the purpose of monitoring the dynamics of nucleosomes in S. cerevisiae strains, the system of...
Oxidative stress in bacteria - with an emphasis on the model organism of Escherichia coli
Moravcová, Andrea ; Lichá, Irena (advisor) ; Čáp, Michal (referee)
Most bacterial species encounter aerobic conditions during their life, which can be toxic. This leads to a state of oxidative stress. Toxicity of aerobic environment results from the chemical properties of molecular oxygen and its reactive species (ROS). Bacteria had to adapt to these conditions in the past to ensure preservation and prosperity. This thesis is preferably focused on oxidative stress adaptations in the most elaborated bacterial model - Escherichia coli. Regulation of adaptations at the regulation of transcription, translation and metabolism level are described with emphasis on molecular mechanisms. The main adaptation mechanism against oxidative stress is the deactivation of ROS, as well as the repair of damaged cell structures (macromolecules). These enzyme activities are regulated by several transcriptional regulators. The transcriptional regulators OxyR and SoxRS have been well studied in E. coli. Even though these regulators are conserved across the bacterial spectrum, they may not have the same function in all organisms. For this reason, also other, more or less studied bacterial species - Bacillus subtilis, Streptomyces coelicolor, Pseudomonas putida, Pseudomonas aeruginosa - were included in this thesis. The various strategies of how these bacteria use not only OxyR and SoxRS...
CRISPR/Cas9-based genome editing in mice: state of the art and future perspectives
Eliáš, Jan ; Kašpárek, Petr (advisor) ; Čáp, Michal (referee)
Mutant mice are crucial tools for understanding gene functions in vivo. Recently, generation of mouse mutants was revolutionized by rapid developement of programmable nucleases, predominantly by the CRISPR/Cas9 system. Genome editing based on introduction of CRISPR/Cas9 components into early stage mouse embyros allows fast and inexpensive generation of gene-deficient animal models, especially when compared to the traditional techniques based on modification of embryonic stem cells (ESCs). The ability of CRISPR/Cas9 to induce double-strand break (DSB) at a given location of genomic DNA enables effective gene-ablation by random modification of the coding sequences or by complete ablation of the gene. However, precise modification of the gene sequences, such as incorporation of a DNA fragment into specific loci, are still difficult to make. In this work, I present a review of CRISPR/Cas9 system, its use in production of mutant mice and possible modifications of the system to increase the efficiency of precise gene-targeting. Keywords: CRISPR/Cas9, mouse, transgenesis, homologous recombination

National Repository of Grey Literature : 44 records found   beginprevious15 - 24nextend  jump to record:
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