Komplementace dsbA mutantního kmene Francisella tularensis a analýza potenciálních vazebných partnerů proteinu DsbA

• Main Author: Šenitková, Iva
• Other Authors: Červený, Lukáš
• Format: Dissertation
• Language: Czech
• Published: 2010
• Online Access: http://www.nusl.cz/ntk/nusl-282893

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Science Candidate: Bc. Iva Šenitková Supervisor: doc. PharmDr. Tomáš Šimůnek, Ph.D. Supervisor - consultant: RNDr. Petra Špidlová, Ph.D. Title of diploma thesis: Complementation of Francisella tularensis dsbA mutant strain and analysis of potential binding partners of DsbA protein Conserved hypothetical lipoprotein FTT1103 is a virulence factor of gram- negative intracellular bacterium F. tularensis. This protein shares homology with proteins of disulfide oxidoreductase DsbA family. These proteins catalyse formation of disulfide bonds, which are essential for forming proteins conformation, for activity and stability of proteins. The aim of this study was to identify binding partners of lipoprotein FTT1103, commonly known as DsbA. The knowledge of these partners could help us in understanding a role of lipoprotein DsbA in virulence F. tularensis. We prepared fused ftt1103 gene with sequence coding FLAG® tag and cloned this fused gene into plasmid vector which can be replicated in F. tularensis. We used electroporation for introducing plasmid vector into the in frame deletion mutant F. tularensis subsp. holarctica strain FSC200/∆1103 thereby we complemented this deletion mutant in trans. Imunoprecipitation by...