Isolation of chimaeric forms of elongation factor EF-Tu by affinity chromatography.
Six different recombinant chimaeric forms of a three-domain protein, proteosynthetic elongation factor Tu (EF-Tu), composed of domains of EF-Tu of mesophilic (Escherichia coli) and thermophilic (Bacillus stearothermophilus) origin as well as free N-terminal domains of EF-Tu, and the whole recombinant EF-Tus of both organisms were prepared and isolated by the GST (glutathione S-transferase) fusion technology. Several modifications in the standard isolation and purification procedures are described that proved necessary to obtain the proteins in a purified and undegraded form.
Tomincová
Hana
H
Department of Protein Biosynthesis, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague.
Krásný
Libor
L
Jonák
Jirí
J
D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D010444:Peptide Elongation Factor Tu / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*
D011993:Recombinant Fusion Proteins; D010444:Peptide Elongation Factor Tu