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Effect of knock out of yxkO gene on environmental stress adaptation in genus Bacillus
Tkadlec, Jan ; Lichá, Irena (advisor) ; Krásný, Libor (referee)
We have previously characterized a Bacillus subtilis mutant defective in growth and osmoadaptation under limited K+ concentrations. In this mutant, the yxkO gene encoding a putative ribokinase is disrupted. This gene is supposed to belong to the sigma B operon and its expression is induced after osmotic, heat and ethanol shock. In comparison to the wild type, this mutation causes pleiotropic changes in host phenotype. In addition to its osmosensitivity, the mutant differs in cell shape, motility and ability to produce endospores. Our goal was to focus on manifestations of the mutation in the yxkO gene in other bacteria of the genus Bacillus. Using plasmid pMUTIN4 we have prepared mutants with disruptions of this gene derived from Bacillus amyloliquefaciens and Bacillus subtilis subsp. spizizenii strains differing in the yxkO surroundings and in the level of laboratory domestication. As in the previous study (with laboratory strain Bacillus subtilis 168) we demonstrate impaired ability of the mutant strain derived from Bacillus amyloliquefaciens to grow in potassium limitation and osmotic shock. We have studied this phenomenon at the level of the growth dynamics of the bacterial culture. We have also detected an increased sensitivity of the strain derived from Bacillus amyloliquefaciens to...
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General stress response in Staphylococcus aureus and its role in adaptation to hyperosmotic stress
Kotková, Hana ; Lichá, Irena (advisor) ; Seydlová, Gabriela (referee)
Hyperosmotic stress induces a stress reponse in a halotolerant bacteria Staphylococcus aureus that helps it to survive even the osmotic conditions that are inhibiting for most of the bacteria. Stress responses of S. aureus include a general stress response that activates rapidly on a wide range of stresses and is ensured by expression of genes induced by an alternative sigma B subunit of RNA polymerase (SigB), and more specific responses to hyperosmotic stress associated with transporters of potassium and transporters of compatible solutes. Also, some virulence factors (such as the adhesive Ebh protein) allow S. aureus to survive hyperosmotic stress within the host, thereby helping to colonize it. It is currently known that the general stress response directly and indirectly participates in the specific stress adaptation through the Kdp transport system for potassium (Kdp transporter) and transporters of compatible solutes (PutP and OpuD). Expression of the Kdp transporter and the large S. aureus Ebh protein is SigB dependent, via Agr or ArlRS virulence regulator systems. The formation of a biofilm, in which SigB is significantly involved, may also enable to colonize host niches with low water activity. Hyperosmotic stress by triggering SigB initiates a complex response that is associated not only...
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Effect of knock out of yxkO gene on environmental stress adaptation in genus Bacillus
Tkadlec, Jan ; Lichá, Irena (advisor) ; Krásný, Libor (referee)
We have previously characterized a Bacillus subtilis mutant defective in growth and osmoadaptation under limited K+ concentrations. In this mutant, the yxkO gene encoding a putative ribokinase is disrupted. This gene is supposed to belong to the sigma B operon and its expression is induced after osmotic, heat and ethanol shock. In comparison to the wild type, this mutation causes pleiotropic changes in host phenotype. In addition to its osmosensitivity, the mutant differs in cell shape, motility and ability to produce endospores. Our goal was to focus on manifestations of the mutation in the yxkO gene in other bacteria of the genus Bacillus. Using plasmid pMUTIN4 we have prepared mutants with disruptions of this gene derived from Bacillus amyloliquefaciens and Bacillus subtilis subsp. spizizenii strains differing in the yxkO surroundings and in the level of laboratory domestication. As in the previous study (with laboratory strain Bacillus subtilis 168) we demonstrate impaired ability of the mutant strain derived from Bacillus amyloliquefaciens to grow in potassium limitation and osmotic shock. We have studied this phenomenon at the level of the growth dynamics of the bacterial culture. We have also detected an increased sensitivity of the strain derived from Bacillus amyloliquefaciens to...
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