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National Repository of Grey Literature 2 records found  Search took 0.01 seconds. 
Production and purification of recombinant human xenobiotic nuclear receptors
 Menclová, Alena ; Wsól, Vladimír (advisor) ; Pávek, Petr (referee)
The submitted thesis deals with the production and purification of recombinant human xenobiotic nuclear receptors (NRs). This study was focused on the production of ligand - binding domains (LBDs) of human constitutive androstane receptor (hCAR) and human pregnane X receptor (hPXR). These receptors belong to the subfamily NR1I NRs and serve as key regulators of drugs metabolism. They are also implicated in many human diseases such as inflammatory bowel disease or diabetes and obesity. The cDNA of recombinant proteins, which serves for production of hCAR and hPXR LBDs, was cloned into bacterial expression vector pET-15b (Novagen). The gained constructs were propagated in E. coli cells of DH5 strain. The isolated plasmids were analyzed after colony PCR and restriction analysis for the correct insert on agarose gels. The clones with correct inserts were subsequently transformed into E. coli cells strain BL21 DE3 and recombinant protein was synthesized in expression system, which was regulated by concentration of isopropyl-1-thio--D-galactopyranosid (IPTG) under optimal conditions. Production of hCAR and hPXR LBD was observed in defined concentrations of IPTG, temperatures and cell densities (OD600). The results revealed the following optimal conditions for protein production; for the hCAR LBD it was...
Detailed record
Production and purification of recombinant human xenobiotic nuclear receptors
 Menclová, Alena ; Pávek, Petr (referee) ; Wsól, Vladimír (advisor)
The submitted thesis deals with the production and purification of recombinant human xenobiotic nuclear receptors (NRs). This study was focused on the production of ligand - binding domains (LBDs) of human constitutive androstane receptor (hCAR) and human pregnane X receptor (hPXR). These receptors belong to the subfamily NR1I NRs and serve as key regulators of drugs metabolism. They are also implicated in many human diseases such as inflammatory bowel disease or diabetes and obesity. The cDNA of recombinant proteins, which serves for production of hCAR and hPXR LBDs, was cloned into bacterial expression vector pET-15b (Novagen). The gained constructs were propagated in E. coli cells of DH5 strain. The isolated plasmids were analyzed after colony PCR and restriction analysis for the correct insert on agarose gels. The clones with correct inserts were subsequently transformed into E. coli cells strain BL21 DE3 and recombinant protein was synthesized in expression system, which was regulated by concentration of isopropyl-1-thio--D-galactopyranosid (IPTG) under optimal conditions. Production of hCAR and hPXR LBD was observed in defined concentrations of IPTG, temperatures and cell densities (OD600). The results revealed the following optimal conditions for protein production; for the hCAR LBD it was...
Detailed record

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1 Menclová, Anika
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