Construction of unmarked mutants in gene coding for Ser/Thr protein kinases and phosphoprotein phosphatase of Pseudomonas aeruginosa and analysis of theur properties

0100410 - MBU-M 20040099 RIV CZ eng C - Conference Paper (international conference)
Nedvědová, Jana - Lněnička, Petr - Hercík, Kamil - Branny, Pavel
Construction of unmarked mutants in gene coding for Ser/Thr protein kinases and phosphoprotein phosphatase of Pseudomonas aeruginosa and analysis of theur properties.
[Konstrukce neoznačených mutantů v genech kódujících Ser/Thr proteinkinasy a fosfoproteinfosfatasu Pseudomonas aeruginosa a analýza jejich vlastností.]
European Congress of Clinical Microbiology and Infectious Diseases /14./. Praha, 2004, s. 730.
[European Congress of Clinical Microbiology and Infectious Diseases /14./. Praha (CZ), 01.05.2004-04.05.2004]
Institutional research plan: CEZ:AV0Z5020903
Keywords : flp-frt * dna
Subject RIV: EE - Microbiology, Virology

To investigate the importance of individual protein kinases and phosphatases in P. aeruginosa physiology we prepared the single and double mutants in the coresponding genes. Single mutants in protein kinase genes stk1, ppkB and protein phosphatase gene stp1 were prepared by the method of gene replacement with subsequent excision of antibiotic cassettes using Flp-FRT recombination system. Two-dimensional gel electrophoresis was used to analyze the patterns of phosphoproteins obtained in in vitro protein kinase assays with the cell-free lysates of wild type as well as all mutant strains.

Abychom zjistili význam jednotlivých proteinkinas a proteinfosfatas Pseudomonas aeruginosa byly připraveny mutanty v jednotlivých genech. Jednoduché mutanty byly připraveny metodou „gene replacement“ a rezistenční kazety byly posléze vyštěpeny pomocí Flp-FRT rekombinačního systému. Pomocí dvourozměrné elektroforézy byly porovnány fosforylační profily divokého typu a mutantů získané in vitro.
Permanent Link: http://hdl.handle.net/11104/0007912