Počet záznamů: 1
Focused directed evolution of beta-glucosidases: theoretical versus real effectiveness of a minimal working setup and simple robust screening
- 1. 0440786 - BFU-R 2015 RIV NL eng J - Článek v odborném periodiku
Mazura, P. - Filipi, T. - Souček, P. - Brzobohatý, Břetislav
Focused directed evolution of beta-glucosidases: theoretical versus real effectiveness of a minimal working setup and simple robust screening.
Carbohydrate Research. Roč. 346, č. 2 (2011), s. 238-242. ISSN 0008-6215
Institucionální podpora: RVO:68081707
Klíčová slova: Directed evolution * beta-Glucosidase * Mutagenesis
Kód oboru RIV: BO - Biofyzika
Impakt faktor: 2.332, rok: 2011
Here we present an optimized procedure to generate amino acid variations at specific site(s) of proteins, followed by a simple one-step screen for mutants with the desired beta-glucosidase activity. The procedure was evaluated by introducing sequence variation into a codon specifying a non-functional variant of the catalytic nucleophile (E401) of the maize beta-glucosidase Zm-p60.1. Observed and theoretically expected frequencies of the four possible variants of the codon and the two possible phenotypes (functional and non-functional) were investigated. Deviations in codon and phenotype frequencies were expressed as a coefficient. This coefficient was then used to estimate the extent of over-sampling, of the mutant library, which would be necessary to compensate for the underrepresentation of some sequences.
Trvalý link: http://hdl.handle.net/11104/0243905