|
|
Study of effect of Bacillus subtilis yxkO gene on motility during stress response to osmotic upshift.
Streitová, Eliška ; Lichá, Irena (advisor) ; Krásný, Libor (referee)
Bacillus subtilis is gram-positive soil bacteria. In its natural environment it is constantly exposed to changes of chemical and physical conditons, including changes of osmolality. It responds to high osmolality by transporting of potassium ions and afterthat transporting and/or synthetising of compatible solutes. In last years the mutant strain Bacillus subtilis L-42 was isolated with non-specific insertional mutagenesis (mini Tn10) in our laboratory. This strain displays limited growth and inability to cope with hyperosmotic shock in a defined medium with potassium concentration of < 1 mmol/l. Insertion of transposon was located in yxkO gene which encodes a protein of unknown biological function. Some other data also indicate a possible role of disruption of yxkO gene in regulation of expression of hag gene, which encodes flagelin - a pivotal protein of bacterial flagellum. The goal of this thesis was to clarify if the disruption of yxkO gene influences motility and whether is affected the transcription of hag gene. With integrative vector pMUTIN4 a mutant strain with specific mutation of yxkO gene was prepared. Vector was pasted into chromosome of Bacillus subtilis strain 1A839 - genotype of this strain allows to extrude the known transcriptional regulation of hag gene. Cell's motility was...
|
|
|
Isolation of bacterial flagellin as an immunogen for the preparation of antibodies for cystic fibrosis patients
Supová, Veronika ; Hodek, Petr (advisor) ; Kubíčková, Božena (referee)
Cystic fibrosis (CF) is an autosomal recessive disease caused by CFTR gene mutation. The consequence of this mutation is an improper function of the chloride channel consisting of CFTR protein. Due to this abnormality, the transport of chloride ions is either reduced or inhibited completely, which leads to a mucus secretion. Mucus is mostly created in the lungs and it is the ideal environment for pathogenic bacteria like Pseudomonas aeruginosa (PA). Pseudomonas aeruginosa is an aerobic, gram-negative conditioned pathogen occurring in patients with weakened immunity such as patients with CF, who are often hospitalized. PA has one polar flagellum which contains filament composed of a protein called flagellin. The flagellum is one of the most important virulence factors of PA bacteria. This thesis focused on the isolation of flagellin from Pseudomonas aeruginosa flagella. The isolated flagellin will serve as an antigen for the preparation of prophylactic antibodies for CF patients. The isolation of flagellin was carried out by four methods using combinations of precipitation and centrifugation. The published methods were optimized to achieve the isolation of flagellin. Final samples and intermediates were analyzed by SDS-electrophoresis on polyacrylamide gel and the presence of flagellin was verified...
|
|
|
Study of effect of Bacillus subtilis yxkO gene on motility during stress response to osmotic upshift.
Streitová, Eliška ; Lichá, Irena (advisor) ; Krásný, Libor (referee)
Bacillus subtilis is gram-positive soil bacteria. In its natural environment it is constantly exposed to changes of chemical and physical conditons, including changes of osmolality. It responds to high osmolality by transporting of potassium ions and afterthat transporting and/or synthetising of compatible solutes. In last years the mutant strain Bacillus subtilis L-42 was isolated with non-specific insertional mutagenesis (mini Tn10) in our laboratory. This strain displays limited growth and inability to cope with hyperosmotic shock in a defined medium with potassium concentration of < 1 mmol/l. Insertion of transposon was located in yxkO gene which encodes a protein of unknown biological function. Some other data also indicate a possible role of disruption of yxkO gene in regulation of expression of hag gene, which encodes flagelin - a pivotal protein of bacterial flagellum. The goal of this thesis was to clarify if the disruption of yxkO gene influences motility and whether is affected the transcription of hag gene. With integrative vector pMUTIN4 a mutant strain with specific mutation of yxkO gene was prepared. Vector was pasted into chromosome of Bacillus subtilis strain 1A839 - genotype of this strain allows to extrude the known transcriptional regulation of hag gene. Cell's motility was...
|
|
| |
|
| |
guest ::
