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Yeast isogenic system as a method for study of IFI16 protein interactions with DNA
Kratochvilová, Libuše ; Šedrlová, Zuzana (referee) ; Brázda, Václav (advisor)
This bachelor thesis deals with the binding of interferon gamma-induced protein 16 (IFI16) to the secondary local structures of the G-quadruplex (G4) and its mutations in the single-hybrid yeast system (Y1H). The IFI16 protein in the cell recognizes its own and foreign or damaged DNA, is involved in the formation of the inflammasome and induces the expression of type I interferon (IFN-I). It is also involved in the regulation of transcription and restriction of viral infection. It has been shown that the IFI16 protein binds preferentially to G-quadruplex structures and is able to stabilize them by this binding. G-quadruplexes are classified as non-canonical DNA and RNA structures formed by G-rich sequences. They are easily formed under physiological conditions and are found in a number of important regulatory structures of the genome such as telomeres or oncogene promoters. They are also part of a number of viral genomes. This makes them excellent potential targets in the treatment of cancer and viral diseases. In the first part of the work, new reporter strains of S. cerevisiae yeasts were prepared by the Delitto Perfetto method, differing only in sequence with the potential for G-quadruplex formation, which was designed and analyzed by the DNA Analyzer program. The correctness of the inserted sequences was verified by PCR and Sanger sequencing and comparison with the supplied oligonucleotide sequences by the Blast program. In the second part of the work, the newly prepared strains were transformed with vectors for the expression of p53, IFI16 proteins, and the effect of IFI16-G4 binding on the expression of the gene in connection with the tumor suppressor p53 was assessed using luciferase reporter assays. The evaluation was performed on the basis of a statistical analysis of the magnitudes of the effects obtained after normalization of the luminescence signal on the optical density of the culture at a wavelength of 600 nm. The results show that the IFI16 protein has a different effect on the trans-activation potential of the p53 tumor suppressor depending on binding to emerging structures near the reporter gene promoter, and that a G4Hunter threshold of at least 1,591 had to be reached and taken into account to successfully form a G-quadruplex the potential of the sequence to successfully form at least 2 G-tetrads.
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Effect of added carbohydrates on the quality of cereal kvass and kvass bread
Stoklásková, Olga ; Vítová, Eva (referee) ; Diviš, Pavel (advisor)
The aim of this bachelor thesis was to characterize selected chemical, senzoric and rheologic properties of sourdough breads with added particular saccharides in the dough. In the theoretical part, processes during mixing, proofing and scaling of the sourdough in fixed conditions are analysed; researched sugars are characterized and their influence on the sourdough product, resulting from present research, is discussed. The experimental part focuses on particular analyses, that influence the quality of sourdough product and determine so its market demand. According to the analysis, sourdough fortification of low honey concentration tends to the maximal volume of the loaf and the most tender crumb at the same time. Higher additions of white table sugar increase the total acidity of the dough the most significantly; unlike, a malt flour provides a smooth, sweetish aroma to the bread. This is because of inhibition of acetic acid production by maltose during sourdough fermentation. Unfortunately, lower concentration of acetate and higher content of remaining sugars in the same time is a factor of shortening the microbial shelf-life of the bread after its baking.
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Srovnání pivovarských a vinných kmenů kvasinek v různých fermentačních podmínkách
Ryšavý, Daniel
The thesis on the topic Comparison of brewer’ and wine’ yeast strains in various fermentation conditions has been prepared at the Department of Postharvest Technology of Horticultural Products at the Faculty of Horticulture, Mendel University in Brno in 2019/2020. The thesis deals with the taxonomy and the properties of yeasts and their useage and focuses mainly on brewer's and wine yeast strains. In the practical part, the analytical parameters of wort fermented by selected brewer's and wine yeast strains have been evaluated and compared. By these yeasts their influence on the sensory quality of the produced beer has also been evaluated.
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Yeast isogenic system as a method for study of IFI16 protein interactions with DNA
Kratochvilová, Libuše ; Šedrlová, Zuzana (referee) ; Brázda, Václav (advisor)
This bachelor thesis deals with the binding of interferon gamma-induced protein 16 (IFI16) to the secondary local structures of the G-quadruplex (G4) and its mutations in the single-hybrid yeast system (Y1H). The IFI16 protein in the cell recognizes its own and foreign or damaged DNA, is involved in the formation of the inflammasome and induces the expression of type I interferon (IFN-I). It is also involved in the regulation of transcription and restriction of viral infection. It has been shown that the IFI16 protein binds preferentially to G-quadruplex structures and is able to stabilize them by this binding. G-quadruplexes are classified as non-canonical DNA and RNA structures formed by G-rich sequences. They are easily formed under physiological conditions and are found in a number of important regulatory structures of the genome such as telomeres or oncogene promoters. They are also part of a number of viral genomes. This makes them excellent potential targets in the treatment of cancer and viral diseases. In the first part of the work, new reporter strains of S. cerevisiae yeasts were prepared by the Delitto Perfetto method, differing only in sequence with the potential for G-quadruplex formation, which was designed and analyzed by the DNA Analyzer program. The correctness of the inserted sequences was verified by PCR and Sanger sequencing and comparison with the supplied oligonucleotide sequences by the Blast program. In the second part of the work, the newly prepared strains were transformed with vectors for the expression of p53, IFI16 proteins, and the effect of IFI16-G4 binding on the expression of the gene in connection with the tumor suppressor p53 was assessed using luciferase reporter assays. The evaluation was performed on the basis of a statistical analysis of the magnitudes of the effects obtained after normalization of the luminescence signal on the optical density of the culture at a wavelength of 600 nm. The results show that the IFI16 protein has a different effect on the trans-activation potential of the p53 tumor suppressor depending on binding to emerging structures near the reporter gene promoter, and that a G4Hunter threshold of at least 1,591 had to be reached and taken into account to successfully form a G-quadruplex the potential of the sequence to successfully form at least 2 G-tetrads.
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Využítí kvasinkových kmenů při výrobě piva
Süss, Jan
This final thesis is focused on topic of use the yeast strains in beer production. In first part of thesis is descripted brief overview of beer making process, its nutritional values, composition and also inputs needed for its production, namely malt, hops, water and yeast. Following part of thesis considers overall characteristics of yeast strains, their reproduction and their importance in food production industry. After general description the thesis is focused on characteristics of individual strains of yeast used in top and bottom fermentation process, which are used in Czech Republic and also in foreign countries. In following point is discussed the characteristic of substances negatively influencing the dynamics and course of fermentation process. Consecutively the undesirable genera of yeasts are described, which could occur at wrong brewer's practice and could negatively alter the qualitative and sensoric properties of beer.
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Vliv kvasinkové mikroflóry na finální jakost vína
Machalínková, Tereza
This thesis examines the influence of yeasts on the final stage of wine, consists of theoretical and practical parts. The theoretical part deals with the yeast themselves, their description, nutrition, the most important yeast for wine fermentation methods and influences that affect the fermentation process. The second part is devoted to work for bridges for optimal fermentation, there must be engaged in expanding and methods of sludge. The next section is devoted to chemical reactions during fermentation, wine composition and in the last part describes the training and maturation of wine. In the practical part is devoted to the production of wine from Pinot Blanc and his subsequent analytical analysis, determination of individual elements such as alcohol, glucose, fructose, glycerol and total acidity. In the last phase is devoted to the evaluation of individual analysis and discussion.
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Identification of yeasts genus Saccharomyces from leaves, grapes and must of grape-vine.
Škodová, Anna ; Štros, Michal (referee) ; Vránová, Dana (advisor)
This diploma thesis is concerned about the PCR-RFLP method for identification of yeasts genus Saccharomyces from leaves, grapes and must of white wine. The thesis describes an obtaining of pure cultures of yeasts, isolation DNA and amplification of specific segments of DNA using PCR method. Splitting of this segments by restriction endonukleases and a detection of final fragments by zone electrophoresis are mentioned too. Length of fragments and numbers of restriction fragments, which are characteristic of every species, enable a determination of microorganisms and their inclusion into the system. The basic informations on yeasts and molecular methods for their identification are named in the theoretical part of the thesis. The main part of the thesis deals with the processing of grapes of grape-wine and the manufacturing of white wine.
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Vliv vybraných preparátů při technologii výroby bílého vína na jeho výslednou kvalitu
Žiška, Zbyněk
This paper assesses the influence of yeast agent on the final quality of wine. It consists of two main sections -- theoretical and practical. The Theoretical section explains where yeast is located, describes strains mportant for wine production, fermentation methods and factors that influence the process of fermentation. Next section is dedicated to the biochemistry of fermentation, constitution of wine and to aromatic substances. The last part describes enzymatic agents used for hydrolisis of pectins and glycosidic binding. The experimental part of the thesis describes production of Chardonnay. There is a description of analytical methods used and evalution of results in the form of a discussion in the concluding section.
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