One step purification of Escherichia coli β-glucuronidase

Kinetic Analysis of β-Galactosidase and β-Glucuronidase Tetramerization Coupled with Protein Translation

“…1). As reported previously (22,23), GAL and GUS both assemble into tetramers, starting from monomer to dimer, followed by dimer to tetramer, which we also showed by size exclusion chromatography (Fig. 4).…”

Kinetic Analysis of β-Galactosidase and β-Glucuronidase Tetramerization Coupled with Protein Translation

“…1). As reported previously (22,23), GAL and GUS both assemble into tetramers, starting from monomer to dimer, followed by dimer to tetramer, which we also showed by size exclusion chromatography (Fig. 4).…”

Purification and Characterization of a β-Glucuronidase Present During Embryogenesis of the Mollusk Pomacea sp.

“…2B). β-glucuronidases have molecular weights between 69 to 300 kDa and monomeric and tetrameric forms are found in mammals and bacteria [15,16,17]. The enzyme found in Pomacea eggs is different from those already described because it was shown to be a dimeric protein with an estimated Mr of 180 kDa, being probably two identical subunits single protein band of ~90 kDa,.…”

Directed evolution of the surface chemistry of the reporter enzyme β-glucuronidase

“…When the library was induced on plates containing the chromogenic GUS substrate, 5-bromo-4-chloro-3-indolyl-b-D-glucuronide (X-gluc), approximately 80% of the colonies were visibly less green than control colonies expressing only the chromosomal gene (see Experimental protocol). b-Glucuronidase functions as a tetramer 16 , so it was likely that many of the mutations in the highly expressed, plasmid-borne library had a dominant negative effect on the function of the chromosomal gene. This did not deter us from utilizing this library for screening experiments, since successive rounds of DNA shuffling should efficiently select against neutral or deleterious mutations 13 .…”

Kinetic Analysis of β-Galactosidase and β-Glucuronidase Tetramerization Coupled with Protein Translation

“…1). As reported previously (22,23), GAL and GUS both assemble into tetramers, starting from monomer to dimer, followed by dimer to tetramer, which we also showed by size exclusion chromatography (Fig. 4).…”

Purification and Characterization of a β-Glucuronidase Present During Embryogenesis of the Mollusk Pomacea sp.

“…2B). β-glucuronidases have molecular weights between 69 to 300 kDa and monomeric and tetrameric forms are found in mammals and bacteria [15,16,17]. The enzyme found in Pomacea eggs is different from those already described because it was shown to be a dimeric protein with an estimated Mr of 180 kDa, being probably two identical subunits single protein band of ~90 kDa,.…”

Directed evolution of the surface chemistry of the reporter enzyme β-glucuronidase

“…When the library was induced on plates containing the chromogenic GUS substrate, 5-bromo-4-chloro-3-indolyl-b-D-glucuronide (X-gluc), approximately 80% of the colonies were visibly less green than control colonies expressing only the chromosomal gene (see Experimental protocol). b-Glucuronidase functions as a tetramer 16 , so it was likely that many of the mutations in the highly expressed, plasmid-borne library had a dominant negative effect on the function of the chromosomal gene. This did not deter us from utilizing this library for screening experiments, since successive rounds of DNA shuffling should efficiently select against neutral or deleterious mutations 13 .…”

Kinetic Analysis of β-Galactosidase and β-Glucuronidase Tetramerization Coupled with Protein Translation

“…1). As reported previously (22,23), GAL and GUS both assemble into tetramers, starting from monomer to dimer, followed by dimer to tetramer, which we also showed by size exclusion chromatography (Fig. 4).…”

Purification and Characterization of a β-Glucuronidase Present During Embryogenesis of the Mollusk Pomacea sp.

“…2B). β-glucuronidases have molecular weights between 69 to 300 kDa and monomeric and tetrameric forms are found in mammals and bacteria [15,16,17]. The enzyme found in Pomacea eggs is different from those already described because it was shown to be a dimeric protein with an estimated Mr of 180 kDa, being probably two identical subunits single protein band of ~90 kDa,.…”

Directed evolution of the surface chemistry of the reporter enzyme β-glucuronidase

“…When the library was induced on plates containing the chromogenic GUS substrate, 5-bromo-4-chloro-3-indolyl-b-D-glucuronide (X-gluc), approximately 80% of the colonies were visibly less green than control colonies expressing only the chromosomal gene (see Experimental protocol). b-Glucuronidase functions as a tetramer 16 , so it was likely that many of the mutations in the highly expressed, plasmid-borne library had a dominant negative effect on the function of the chromosomal gene. This did not deter us from utilizing this library for screening experiments, since successive rounds of DNA shuffling should efficiently select against neutral or deleterious mutations 13 .…”