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Construction of vectors for heterologous expression of human cytochrome P450 1B1
Sojka, Pavel ; Martínek, Václav (advisor) ; Moserová, Michaela (referee)
The thesis was worked out in Laboratory of Molecular Carcinogenesis and Drug Development, which is focus on study of drug metabolizing enzymes including cytochromes P450. Cytochromes P450 are participating at initial phase of biotransformation of xenobiotics and endogenous substances and metabolism of several endogenous compounds, i.e. steroids. This work is focused on construction of expression vectors, based on the plasmid pET- 22b, suitable for heterologous expression of the human cytochrome P450 1B1. This enzyme is predominantly present in the endoplasmic reticulum of extra-hepatic tissues and its expression is induced by dioxins and polycyclic aromatic hydrocarbons. The human gene for cytochrome P450 1B1 was modified using PCR. The cleavage sites for restriction endonucleases were added to both ends of the gene. Another construct also contained N-terminal histidine tag to facilitate easier purification of the enzyme. Both insert and digested plasmids were verified using the agarose electrophoresis and used for ligation and transformation into competent cells (E. coli DH5. Final steps in construction was, however, not successful, probably due to low yields of DNA fragment extraction from agarose gels. Key words: cytochrome P450 1B1, carcinogenesis, plasmid, heterologous expression [In Czech]
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Language of Primary School Pupils
Cmíralová, Hana ; Janovec, Ladislav (advisor) ; Sojka, Pavel (referee)
This work deals with contemporary speech elementary school pupils. The theoretical part of the author accesses the current terminology of unstructured language services, defines the term speech of students in relation to the slang and presents the basic characteristics of speech of pupils and their holders. The material from witch the author builds vocabulary to describe student speech, was acquired through long-term academic research environment. Based on the examination of the recovered materials, the author of several significant conclusions concerning the speech of students. Speech students include specific language statement, which wants pupils to show their unique relationship with the school and the world around them. New words, they create, are a reflection of how the school and, upon it, are experiencing. For the speech of students is typical naming considerable ingenuity, applied in a number of her naming practices. In addition to the traditional derivation is a common distortion and mechanical reduction. Many is the transfer of meaning, especially metaphorical and acceptance of foreign languages. Student vocabulary is highly variable and includes a large number of new and traditional names, but names and fhrases are absolete or do not. This variability pupil vocabulary derives mainly from...
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Heterologous expression and isolation of human cytochrome P450 1B1
Sojka, Pavel ; Martínek, Václav (advisor) ; Moserová, Michaela (referee)
Cytochromes P450 are heme enzymes with very broad substrate specificity, they can oxidize tens to hundreds of different substrates including both eobiotics and xenobiotics, but with varying efficiencies and kinetics. They are responsible for the metabolic activation of many carcinogens resulting in formation of reactive intermediates, these reactive species participate in the formation of DNA adducts and also increase of oxidative stress. Eukaryotic cytochromes P450 are membrane bound enzymes found mostly in the endoplasmic reticulum. In vertebrates, they are expressed in a variety of tissues mostly in the liver, but also in kidney, lung, skin and others. The cytochrome P450 1B1 is an inducible enzyme which occurs mainly in the lung and skin. Its expression is induced predominantly by the presence of polycyclic aromatic hydrocarbons, dioxins and heterocyclic amines. The human cytochromes P450 are typically obtained using heterologous expression and isolated as a C-terminally modified hexa-histaq constructs using immobilized metal affinity chromatography. This thesis focuses on effect of C-terminal modifications on activity of human cytochrome P450 1B1. First, the two expression vectors encoding the human form of cytochrome P450 1B1 were prepared, one contained a classical C-terminal hexa-histaq...
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The Language of Partly Prepared Speech (guides)
Hlaváčková, Klára ; Janovec, Ladislav (advisor) ; Sojka, Pavel (referee)
Professional public show considerable interest in a form of spoken language. The dissertation The Language of Semi-prepared Speeches (Guides in Castles and Chateaux)' has the honour to extend this area of research. The base of this essay is material obtained by field research. Speeches of ten guides in twelve sights were screened by phonetic and phonological analysis. The aim of this analysis was a description of the language in the communication that springs up between visitors of sights and their guides. Furthermore, we are dealing with basic anomalies of literary pronunciation and we are trying to observe some stereotypes in the speech of this profession group. Step by step this dissertation is dealing with vowels, it is observing their quantity (stretching, reducing), quality (open, closed pronunciation, reduction, vowels changes, elision) and prothetic form. Chapters about analysis of consonants, especially their elision, are the next. There is also a passage analysing sentence intonation. The end is dealing with a speech tempo. What is the speech of the surveyed guides like then? The answer is in the content of the dissertation.
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Morphological Norms in the Radio Programme Interview Plus
Stupková, Nikola ; Sojka, Pavel (advisor) ; Adam, Robert (referee)
The diploma thesis aims to map the use of certain morphological phenomena by the hosts as well as the guests of Interview Plus broadcast at Český rozhlas Plus. The thesis focuses on morphological norms, in particular on three morphological phenomena - competitiveness of conditional forms "bychom", "bysme", the concord in plural neuters in the nominative, accusative, and instrumental cases. The aforementioned phenomena have been chosen because they are problematic, and their nonstandard usage has been infiltrating standard utterances. Both standard and nonstandard forms of all phenomena have been detected. The findings have been interpreted from different perspectives. Firstly, social factors, such as the speakers' profession or their gender, were taken into consideration. Secondly, the thesis observes the proportion of standard to nonstandard forms, that is how many speakers use only standard forms, how many speakers use the nonstandard form only, and how many speakers combine the two. Several hypotheses have been formulated. The first hypothesis claims that the hosts use more standard than nonstandard forms. The second hypothesis states the guests use more nonstandard forms than the hosts. The third hypothesis applies only to journalists for it proposes that the journalists use more standard forms...
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Sound properties of moderators' speeches in selected TV talk shows
Baptistová, Lucie ; Vlčková, Jana (advisor) ; Sojka, Pavel (referee)
The thesis investigates speech sound characteristics of three different television talk show hosts. Using the experimental analysis it tries to inspect all their typical attributes, especially the suprasegmental features of their speeches. The selected television hosts are both from public and commercial television stations. The first part of the theoretical section introduces the talk show television programming genre and specifies its broadcasting share among the observed television channels, next the diversity of Czech talk shows is introduced together with the individual hosts whose utterances will be analyzed, also the role of communication in mass media as well as the host journalist figure is presented, and the features of publicly spoken speeches are described. Within the part devoted to languages and speeches the linguistic discipline of phonetics is introduced followed by the individual continuous speech sound properties and their specific attributes. The practical section researches performance of the particular hosts and it specifically tries to deal with their sound aspects in order to evaluate whether some typical attributes can be found (primarily within the areas of intonation, pauzation and tempo). Further it observes whether some same sound attributes are shared among the talk...
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Language of semi-prepared speech
Sojka, Pavel ; Janovec, Ladislav (advisor)
Cílem této práce je hláskoslovná a morfologická analýza mluvených textů rozhlasové publicistiky. Při analýze jsme pracovali se třinácti zaznamenanými a přepsanými texty z let 2002-2005. Především jsme se zabývali otázkou, do jaké míry se v tomto typu projevů dodržuje spisovná norma, a výsledky jsme porovnali se závěry pěti dřívějších výzkumů mluvené češtiny. Zjistili jsme, že výskyt nespisovných prvků v našem vzorku je většinou nižší než v dřívějších výzkumech mluvené češtiny. Zároveň se však potvrdilo, že zejména některé nespisovné prvky mají tendenci pronikat i do projevů zřetelně tíhnoucích ke spisovnosti. Analyzované polopřipravené projevy proto hodnotíme jako projevy jazykově smíšené s převahou spisovných prvků.
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