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Isolation of chimaeric forms of elongation factor EF-Tu by affinity chromatography
Hana Tomincová1, Libor Krásný, Jirí Jonák
1Department of Protein Biosynthesis, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague.
Abstract:
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.