One-step purification of E. coli elongation factor Tu
Knudsen, C.R.; Clark, B.F.; Degn, B.; Wiborg, O.
Biochemistry International 28(2): 353-362
1992
ISSN/ISBN: 0158-5231 PMID: 1456956 Document Number: 388263
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.