Konstrukce neoznačených mutantů v genech kódujících Ser/Thr proteinkinasy a fosfoproteinfosfatasu Pseudomonas aeruginosa a analýza jejich vlastností

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.