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- W4968076 abstract "Directed evolution is widely used in protein engineering forproducing proteins with altered or improved properties. Thispowerful tool includes several main approaches. The first is arandom mutagenesis, which is used for creating random mutationsanywhere in the gene of interest. The second is a targetedrandom mutagenesis, which is utilized for preparing mutantswith random base substitutions in selected hotspots. Inaddition, a site directed mutagenesis is based on rationaldesign and it serves for creating specific amino acidsubstitutions in selected hotspots. In our work two technicalapproaches - the targeted-random mutagenesis via mixture ofoligonucleotide primers and the site directed mutagenesis viatwo primers carrying specific substitution – are employed.Lectins are proteins produced by different organisms. They areable to bind various sugars specifically. This fact is veryimportant especially in host pathogen interactions wherelectins help to pathogen in adhesion to sugar moietiespresented on host cells. Pseudomonas aeruginosa, Ralstoniasolanacearum and Burkholderia cenocepacia are opportunisticpathogens which attack mainly imunocompromised patients. Thesebacteria produce lectins that display significant similarities.The PA-IIL lectin from bacterium P. aeruginosa was chosen as aprimary target gene to be modified via targeted randommutagenesis. This soluble protein is well-characterized and itshows very high affinity for L fucose but it can bind severalmonosaccharides with similar stereochemistry e.g. L-galactoseor D mannose. Orthologues of this lectin have been identifiedin bacteria R. solanacearum (RS IIL) and B. cenocepacia (BC2LA). However, these two PA IIL-like lectins prefer D mannoseinstead of L fucose. The aim of this work is to produceCa-dependent lectins with a sharp specificity to L-fucose. Thelectins with improved properties could be further used inbiotechnology and for bioanalytical purposes." @default.
- W4968076 created "2016-06-24" @default.
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- W4968076 date "2011-01-11" @default.
- W4968076 modified "2023-10-08" @default.
- W4968076 title "Directed evolution of lectins from pathogenic bacteria" @default.
- W4968076 hasPublicationYear "2011" @default.
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