Single-step purification of bacterially expressed polypeptides containing an oligo-histidine domain

Plasmid expression vectors have been constructed that direct the synthesis in Escherichia coli of fusion proteins containing a stretch of six histidine residues at either the N or C terminus. This oligo-histidine domain allows the single-step purification of the fusion proteins, under nondenaturing...

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Bibliographic Details
Published in:	Gene Vol. 111; no. 1; p. 99
Main Authors:	Van Dyke, M W, Sirito, M, Sawadogo, M
Format:	Journal Article
Language:	English
Published:	Netherlands 01-02-1992
Subjects:	Amino Acid Sequence Bacterial Proteins - chemistry Bacterial Proteins - genetics Bacterial Proteins - isolation & purification Base Sequence Chromatography, Affinity Cloning, Molecular DNA, Bacterial Escherichia coli - genetics Genetic Vectors Histidine - genetics Molecular Sequence Data Plasmids Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - isolation & purification
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Summary:	Plasmid expression vectors have been constructed that direct the synthesis in Escherichia coli of fusion proteins containing a stretch of six histidine residues at either the N or C terminus. This oligo-histidine domain allows the single-step purification of the fusion proteins, under nondenaturing conditions, by immobilized metal affinity chromatography on Ni2+ bound to iminodiacetic acid-agarose. Several eukaryotic transcription factors (e.g., the upstream stimulatory factor for the adenovirus major late promoter) have been successfully purified, in an active state, by this method.
ISSN:	0378-1119
DOI:	10.1016/0378-1119(92)90608-R