Production and purification of Japanese quail ovalbumin as fusion protein with glutathione S-transferase in Escherichia coli.
A plasmid encoding a fusion protein interlinked by thrombin recognition sequence between glutathione S-transferase and Japanese quail ovalbumin (without 40 amino acid residues from the 5'-end of the ORF) has been constructed, employing the expression system pGEX-2T. The deglycosylated fusion protein (64 kDa) was purified by affinity chromatography on glutathione agarose beads, analyzed by SDS-polyacrylamide gel electrophoresis, immunochemically detected with antiserum raised against Japanese quail ovalbumin and tested for its stability.
Visvaderová
J
J
Laboratory for Genetic Engineering, Slovak Academy of Sciences, Bratislava.
Albert
S
S
Kosová
A
A
Klaudiny
J
J
Simúth
J
J
D000818:Animals; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D003370:Coturnix; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D006031:Glycosylation; D010047:Ovalbumin / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D010047:Ovalbumin; D005982:Glutathione Transferase