One-step purification of E. coli elongation factor Tu.
The tuf A gene, encoding the E. coli elongation factor Tu, was cloned in the pGEX gene fusion system. Upon expression EF-Tu is fused to glutathione-S-transferase serving as a purification handle with affinity for glutathione immobilised on agarose. This allows purification of EF-Tu in a one-step procedure. The construct was designed in order to make possible the release of authentic EF-Tu by cleaving the fusion protein with the protease factor Xa.
Knudsen
C R
CR
Department of Chemistry, Aarhus University, Denmark.
Clark
B F
BF
Degn
B
B
Wiborg
O
O
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D015951:Factor Xa / Q000378:metabolism; D015870:Gene Expression; D005982:Glutathione Transferase / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010444:Peptide Elongation Factor Tu / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011728:Pyridones / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification*
D000900:Anti-Bacterial Agents; D011728:Pyridones; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D015951:Factor Xa; D010444:Peptide Elongation Factor Tu; C008536:mocimycin