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National Repository of Grey Literature 2 records found  Search took 0.00 seconds. 
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...
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Vliv cílených mutací receptoru cytokininů CRE1/AHK4 z Arabidopsis thaliana na jeho odezvu vůči přirozeným a syntetickým cytokininům
 Horáčková, Kamila
The Arabidopsis thaliana cytokinin receptor CRE1/AHK4 is a part of the signalling pathway by which cytokinin regulates the transcription of target genes via transcription factors. To further elucidate its molecular function, a transgenic E. coli strain KMI001 baring a ß-galactosidase-based reporter system can be employed. The theoretical part of this thesis describes the AHK4 receptor, including its structure, function and previously characterized mutations, and outlines the bacterial reporter system for AHK4 activity assays. The experimental part summarizes the results of ß-galactosidase activity assay optimization, PCR mutagenesis, the transformation of mutagenized plasmids into the E. coli strain KMI001 and receptor activity assays. In accordance with previous reports, the receptor mutant forms exhibited a higher sensitivity to trans-zeatin than the corresponding AHK4 wild-type. Further, the V471A mutant showed a significant decrease in the half maximal effective concentration for 6-benzylaminopurine and thidiazuron.
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See also: similar author names
2 Horáčková, Karolína
6 Horáčková, Kateřina
5 Horáčková, Klára
4 Horáčková, Kristýna
2 Horáčková, Květa
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