Phosphoglucosamine mutase (GlmM), an enzyme taking part in biosynthesis of cell wall, has been recently proven to be essential for Streptococcus pneumoniae. The main goal of this thesis was to prove in vivo that GlmM serine residues S99 and S101 phosphorylation is essential while the necessity of it was already proven indirectly based on transformation efficiency. For this purpose we have prepared a strain with two copies of the glmM gene - the first one with amino acid changes on monitored serine residues located at native locus; and the second ectopic copy of the wild allele of glmM gene under control of inducible zinc promoter. We have observed morphology, growth, and GlmM expression with and without the presence of an inductor. All the observed parameters show that the cells are not viable without ectopic glmM expression, thus the essential protein GlmM is functional only when phosphorylated on S99 and S101 residues. Further, we have attempted to localize the enzyme in the S. pneumoniae cell. We have fused GlmM with fluorescent marker GFP and by using the florescent microscopy we have proved that GlmM is cytoplasmic protein. Another goal of this thesis was to find an unknown third phosforylation site of the GlmM protein which is dependent on the protein kinase StkP. From in vitro kinase assay and...
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:397458 |
| Date | January 2019 |
| Creators | Mühldorfová, Tereza |
| Contributors | Ulrych, Aleš, Lišková, Petra |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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