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Statements

Subject Item
n2:RIV%2F00216224%3A14310%2F10%3A00044241%21RIV11-GA0-14310___
rdf:type
skos:Concept n15:Vysledek
dcterms:description
Directed evolution is a powerful tool widely used in protein engineering for producing proteins with altered or improved properties. This approach includes a lot of methods used to create diverse mutant libraries and to screen these libraries. In our work two mutation techniques are being used. The first one is random mutagenesis by error-prone PCR (which introduces nucleotide changes during PCR due to the use of error-prone DNA polymerase and reaction conditions). The second one is targeted-random mutagenesis (which employs mixture of special primers carried random base substitutions on selected site). LecB, the gene coding PA-IIL lectin from bacterium Pseudomonas aeruginosa, was chosen as a target gene to be modified via random mutagenesis. This soluble protein is well-characterized and it is indisputable that it displays an unusually high affinity for monosaccharides widely occurring as terminal residues in cell surface glycoconjugates highly present in cystic fibrosis mucins. Directed evolution is a powerful tool widely used in protein engineering for producing proteins with altered or improved properties. This approach includes a lot of methods used to create diverse mutant libraries and to screen these libraries. In our work two mutation techniques are being used. The first one is random mutagenesis by error-prone PCR (which introduces nucleotide changes during PCR due to the use of error-prone DNA polymerase and reaction conditions). The second one is targeted-random mutagenesis (which employs mixture of special primers carried random base substitutions on selected site). LecB, the gene coding PA-IIL lectin from bacterium Pseudomonas aeruginosa, was chosen as a target gene to be modified via random mutagenesis. This soluble protein is well-characterized and it is indisputable that it displays an unusually high affinity for monosaccharides widely occurring as terminal residues in cell surface glycoconjugates highly present in cystic fibrosis mucins.
dcterms:title
Methods of directed evolution of PA-IIL lectin Methods of directed evolution of PA-IIL lectin
skos:prefLabel
Methods of directed evolution of PA-IIL lectin Methods of directed evolution of PA-IIL lectin
skos:notation
RIV/00216224:14310/10:00044241!RIV11-GA0-14310___
n4:aktivita
n16:S n16:P n16:Z
n4:aktivity
P(GA303/09/1168), P(GD301/09/H004), P(LC06030), S, Z(MSM0021622413)
n4:dodaniDat
n6:2011
n4:domaciTvurceVysledku
n17:5522064 n17:6666477 n17:5616697
n4:druhVysledku
n14:O
n4:duvernostUdaju
n18:S
n4:entitaPredkladatele
n12:predkladatel
n4:idSjednocenehoVysledku
270725
n4:idVysledku
RIV/00216224:14310/10:00044241
n4:jazykVysledku
n8:eng
n4:klicovaSlova
directed evolution; random mutagenesis in vitro; targeted-random mutagenesis; lectin PA-IIL; Pseudomonas aeruginosa
n4:klicoveSlovo
n5:lectin%20PA-IIL n5:targeted-random%20mutagenesis n5:Pseudomonas%20aeruginosa n5:directed%20evolution n5:random%20mutagenesis%20in%20vitro
n4:kontrolniKodProRIV
[56C9115063C9]
n4:obor
n9:CE
n4:pocetDomacichTvurcuVysledku
3
n4:pocetTvurcuVysledku
3
n4:projekt
n10:GD301%2F09%2FH004 n10:GA303%2F09%2F1168 n10:LC06030
n4:rokUplatneniVysledku
n6:2010
n4:tvurceVysledku
Pokorná, Martina Wimmerová, Michaela Mrázková, Jana
n4:zamer
n13:MSM0021622413
n7:organizacniJednotka
14310