Phosphoglucosamine mutase (GlmM) is an enzyme of bacterial cell wall biosynthesis. The main aim of this thesis was to find out, whether gene glmM is essential for viability of Streptococcus pneumoniae. Therefore, we prepared merodiploid strain containing two copies of glmM; the genomic gene and ectopic copy under control of zinc inducible promoter. Subsequently, depletion strain was prepared by deletion of genomic copy of glmM. This strain was further used for analysis of viability and phenotype features in the medium containing various concentrations of zinc ions, an inducer of ectopic glmM expression. We found out, that the viability of this strain was strictly dependent on the concentration of inducer and further, that depletion of GlmM resulted in remarkable morphological defects. The rescue of mutant strain was observed after addition of inducer up to the level of the control sample. These results have provided the evidence of glmM essentiality for S. pneumoniae viability. Furthermore, we analyzed, whether phosphorylation of key amino acid residues, S99 and S101, is essential for GlmM functionality. Four different strains were prepared by means of site-directed mutagenesis expressing glmM with substitutions of key serine residues for alanine or glutamic acid. Since deletion of chromosomal locus in...

The diploma thesis explores whether the complementation of infinitives and gerunds, functioning as subject clauses, is different. Expecially it compares syntactic structures, relative length and complexity of the complementations and their roles in functional sentence perspective (whether elements are rather thematic or rhematic etc.). Furthemore it explores whether the form of complementation is affected by the position of infinitive of gerund, it means if there is any difference between the complementations of non-finite verb in initial position and extraposition. The research is based on the relevant academic writing and on authentic instances of gerunds and infinitives from the present sources of literature.

Tato práce se zabývá vývojem efektivních technik pro konečné automaty a jejich aplikace. Zejména se věnujeme konečným automatům použitých pří detekci útoků v síťovém provozu a automatům v rozhodovacích procedurách a verifikaci. V první části práce navrhujeme techniky přibližné redukce nedeterministických automatů, které snižují spotřebu zdrojů v hardwarově akcelerovaném zkoumání obsahu paketů. Druhá část práce je je věnována automatům v rozhodovacích procedurách, zejména slabé monadické logice druhého řádů k následníků (WSkS) a teorie nad řetězci. Navrhujeme novou rozhodovací proceduru pro WS2S založenou na automatových termech, umožňující efektivně prořezávat stavový prostor. Dále studujeme techniky předzpracování WSkS formulí za účelem snížení velikosti konstruovaných automatů. Automaty jsme také aplikovali v rozhodovací proceduře teorie nad řetězci pro efektivní reprezentaci důkazového stromu. V poslední části práce potom navrhujeme optimalizace rank-based komplementace Buchiho automatů, které snižuje počet generovaných stavů během konstrukce komplementu.

The present thesis is concerned with the syntactic, semantic and FSP aspects of ditransitive complementation. All these aspects are discussed not only theoretically, but mainly practically in an analysis of two ditransitive verbs: blame and provide. For the purpose of the present analysis, 200 sentences (100 for each of the analyzed verbs) were excerpted from the British National Corpus. The analyzed verbs enter into two possible sentence structures. The first construction includes a subject and verb as well as a direct object and a prepositional object (SVOdOprep); the alternative construction includes a subject and verb as well as an indirect object and a prepositional object (SVOiOprep). One of the points of analysis is a quantitative formulation of the number of occurrences of each of the respective sentence structures for the analyzed verbs within the excerpted material. Within the ditransitive construction we may sometimes encounter object omission of either of the objects (more commonly the indirect object). The analysis concentrates on the possibilities of object omission within ditransitive constructions with the two analyzed verbs. Part-of-speech representation of both objects is also a matter of analysis; there are altogether four possible part-of-speech patterns depending on whether...

In the search for the mitochondrion of oxymonads, DNA of Monocercomonoides exilis - an oxymonad isolated from the gut of Chinchilla, was isolated and its genome was sequenced. Sequencing resulted in a fairly complete genome which was extensively searched or genes for mitochondrion related proteins, but no reliable candidate for such gene was identified. Even genes for the ISC pathway, which is responsible for Fe-S cluster assembly and considered to be the only essential function of reduced mitochondrion-like organelles (MROs), were absent. Instead, we were able to detect the presence of a SUF pathway which functionally replaced the ISC pathway. Closer examination of the SUF pathway based on heterologous localisation revealed that this pathway localised in the cytosol. In silico analysis showed that SUF genes are highly conserved at the level of secondary and tertiary structure and most catalytic residues and motifs are present in their sequences. The functionality of these proteins was further indirectly confirmed by complementation experiments in Escherichia coli where SUF proteins of M. exilis were able to restore at least partially Fe-S cluster assembly of strains deficient in the SUF and ISC pathways. We also proved by bacterial adenylate cyclase two-hybrid system that SufB and SufC can form...

The objective of the thesis is to describe and compare the modifying uses of two adjectives which can appear both as premodifiers as well as bare postmodifiers, namely available and responsible. Both modifying positions are examined with respect to the textual environment, the complexity of noun phrases and adjective phrases in which the adjectives function, reference and other features in order to outline the patterns in which these two adjectives appear. The data for the analytical part have been extracted from the British National Corpus using the basic query and limiting the search to the written part of the corpus. After manual assessment of the initial samples of 500 concordance lines, 154 concordance lines containing the adjective available and 147 containing the adjective responsible proved to be suitable for the analysis. The final samples of the data are further categorized and analysed concerning the complexity of the noun phrase, premodification, postmodification and complementation of the adjectives, semantic preference of the head nouns, and the motivation for the pre- or postnominal placement of the modifiers. Key words: adjective, adjective phrase, premodification, postmodification, complementation, attributive function, predicative function, postpositive function, bare adjective,...

One of the important tasks of virology and immunology is to explore the species- and cell-barriers preventing virus horizontal transmission and reveal the ways how viruses overcome these barriers and "adapt" to different species. This work is based on a well- established retroviral model - avian Rous sarcoma virus (RSV) and studies virus replication blocks in mammalian cells at both pre- and post-integration level. Interaction of the viral envelope glycoprotein (Env) with a specific cellular receptor mediates virus entry into cells. Although mammalian orthologues of specific chicken receptors do not support RSV entry, it was observed that some RSV strains are able to enter mammalian cells. Several RSV-transformed rodent cells lines were described and analysis of provirus H20- RSV in one these cells lines (hamster H-20 tumor cell line) showed multiple mutations including two crucial amino acid substitutions in different regions of Env. Substitutions D32G and L378S confer virus transmission to hamster, human and also chicken cells lacking the appropriate receptor. Altered conformation of H20-RSV Env is similar to a receptor-primed (activated) state of Env. This observation indicates that virus can circumvent the need of original cell receptor because of spontaneous Env activation caused by single...