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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...
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Flagellin and outer surface proteins from Borrelia burgdorferi are not glycosylated
ŠTĚRBA, Ján
Glycosylation of four proteins from Borrelia burgdorferi s.s. was investigated ? flagellins FlaA, FlaB, and outer surface proteins OspA and OspB. Glycosylation of these four proteins was not proved by any of the used techniques. However, other glycan-staining positive proteins were present in the borrelia samples. These proteins were suggested to originate in the culture medium.
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