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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacterial Proteins*
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Base Sequence
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Cloning, Molecular / methods*
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli / genetics*
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Histidine
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Molecular Sequence Data
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Plasmids
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Promoter Regions, Genetic*
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Pseudomonas putida / genetics*
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Pseudomonas putida / metabolism
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / isolation & purification
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Restriction Mapping
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Transcription Factors / biosynthesis*
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Transcription Factors / genetics
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Transcription Factors / isolation & purification*
Substances
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Bacterial Proteins
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PfrA protein, Pseudomonas putida
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Recombinant Proteins
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Transcription Factors
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Histidine