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6S RNA as a tool for increased production of clinically relevant antibiotics
Kostílek, Štěpán ; Bobek, Jan (advisor) ; Ulrych, Aleš (referee)
Streptomyces belong to the group of soil microorganisms, and they are significant producers of biologically active substances used in a wide range of industries, whether it is medicine, agriculture or biotechnology. It is for this reason that the research of these microorganisms is of interest to a number of research institutions. One of the most important groups of these biologically active substances are antibiotics. A significant part of antibiotics produced in nature is produced precisely by streptomyces, and therefore it is in our interest to understand the mechanisms that influence this production. A wide variety of different regulatory mechanisms controlling antibiotic production have been found so far in streptomyces. One of these regulatory mechanisms is the utilization of scr3559 RNA. Scr3559 RNA is a small, non-coding RNA molecule that regulates antibiotic production at the level of DNA transcription. These small RNA molecules mimic the structure of the target promoter sequence and are able to interact with RNA polymerase. The presence of scr3559 RNA has not been confirmed in streptomyces as of yet, but new scr3559 RNA molecules are still being sought. To find new molecules, in silico methods are used, where the similarity of the scr3559 RNA sequence of S. coelicolor with homologous...
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Induction and detection of senescence in vitro
Kostílek, Štěpán ; Stiborová, Marie (advisor) ; Dračínská, Helena (referee)
Nowadays, despite the constant progress of modern medicine and therapeutic methods, one of the most widespread and most serious problems is cancer. The number of patients with cancer increases every year, and in spite of advances made in recent years in this field of medicine, it is still very difficult to find cure to these diseases. This thesis deals with the induction and detection of senescence on two different human cell lines, one cancer derived and one normal cell line, the human neuroblastoma cell line UKF-NB-4 and the human fibroblast line HDFn. Senescence was induced by different concentrations of hydrogen peroxide solution or by long-term cultivation. Senescent cells were detected by histochemical staining of X-Gal as well as evaluation based on morfological changes of the cells. In the HDFn cell line, senescent cells were detected using X-Gal kit. The number of senescent cells increased proportionaly with the concentration of hydrogen peroxide. In the neuroblastoma line, senescence was not detectable by X-Gal kit, although the cells treated withh hydrogen peroxide exhibited morphological features associated with senescence. Key word: senescence, hydrogen peroxide, fibroblasts, neuroblasts, SA-β-Gal
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