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Controlled specific expression and purification of 6 x His-tagged proteins in Pseudomonas
I Bertani1, G Devescovi, V Venturi
1Bacteriology Group, International Centre for Genetic Engineering and Biotechnology, Area Science Park, Padriciano 99, 34012, Trieste, Italy.
FEMS Microbiology Letters
|September 11, 1999
Abstract:
The 6 x His affinity tags have proved invaluable for the exclusive purification of proteins, in Escherichia coli, of genes cloned in frame with a 6 x His tag and a strong inducible promoter. Here, we demonstrate that the system can be extended to Gram-negative bacteria other than E. coli, by the use of compatible broad-host-range plasmids. As an example, the inducible synthesis and specific purification of the Pseudomonas 6 x His-PfrA siderophore regulatory protein in Pseudomonas putida WCS358 is demonstrated.