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Analysis of essentiality of glmM gene coding for phosphoglucosamine mutase of Streptococcus pneumoniae.
Krupička, Jiří ; Branny, Pavel (advisor) ; Čáp, Michal (referee)
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...
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Populism in Representative Democracy: Jeopardy or Necessary Supplement
Bublík, Vojtěch ; Bíba, Jan (advisor) ; Slačálek, Ondřej (referee)
(in English): Main goal of this diploma is to analyze the relationship between populism and representative democracy. The theoretical part of my work is researching populism as a vague concept and determining its minimal definition and esential constituents (I have declined the possibility that populism is only anti-essentialy formed attitude/phenomenon). In the analytical part I've studied the relationship between populism and representative democracy, and defined drawbacks of this uneasy coexistence. However I believe that representative democracy can't be successfully fulfiled without this phenomenon and I have also found not only positive moments of this relationship - I assume that populism is necessary supplement of representative democracy.
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Populism in Representative Democracy: Jeopardy or Necessary Supplement
Bublík, Vojtěch ; Bíba, Jan (advisor) ; Slačálek, Ondřej (referee)
(in English): Main goal of this diploma is to analyze the relationship between populism and representative democracy. The theoretical part of my work is researching populism as a vague concept and determining its minimal definition and esential constituents (I have declined the possibility that populism is only anti-essentialy formed attitude/phenomenon). In the analytical part I've studied the relationship between populism and representative democracy, and defined drawbacks of this uneasy coexistence. However I believe that representative democracy can't be successfully fulfiled without this phenomenon and I have also found not only positive moments of this relationship - I assume that populism is necessary supplement of representative democracy.
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Analysis of essentiality of glmM gene coding for phosphoglucosamine mutase of Streptococcus pneumoniae.
Krupička, Jiří ; Branny, Pavel (advisor) ; Čáp, Michal (referee)
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...
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