Optimalizace exprese vybrané lidské karbonylreduktasy v Escherichia coli

• Main Author: Ježková, Veronika
• Other Authors: Novotná, Eva
• Format: Dissertation
• Language: Czech
• Published: 2016
• Online Access: http://www.nusl.cz/ntk/nusl-345291

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Veronika Ježková Supervisor: RNDr. Eva Novotná, PhD. Title: The optimization of expression of the selected human carbonyl reductases in Escherichia coli The present thesis called The optimization of expression of the selected human carbonyl reductases in Escherichia coli is divided into two parts. The first one is theoretical and the second one is experimental. The theoretical part presents a brief information summary about selected enzymes: carbonyl reductases 1 (CBR1), quinone oxidoreductase 1 (NQO1) and DHRS2 (Hep27). It also includes a theoretical summary of laboratory methods which were used in the experimental part. The experimental part aims to optimize the express conditions of the selected enzymes in the bacterial expression system Lemo21(DE3). The plasmid pET-28b(+), which encodes the selected enzymes, is transformed by a thermal shock into competent bacterial cells Lemo21(DE3). In these cells a protein expression was carried out at different concentrations of L-rhamnose. We also purified the expressed recombinant protein NQO1 by the affinity chromatography method. The success rate of protein expression was examined by SDS-PAGE or Western Blot. Based on the obtained results...