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

Michael W. Van Dyke; Mario Sirito; Michèle Sawadogo

doi:10.1016/0378-1119(92)90608-R

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

Michael W. Van Dyke, Mario Sirito, Michèle Sawadogo

Genetics

Research output: Contribution to journal › Article › peer-review

138 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	99-104
Number of pages	6
Journal	Gene
Volume	111
Issue number	1
DOIs	https://doi.org/10.1016/0378-1119(92)90608-R
State	Published - Feb 1 1992

Keywords

Affinity chromatography
Escherichia coli
expression vector
iminodiacetic acid-agarose
plasmid pUC19
recombinant DNA

ASJC Scopus subject areas

Genetics

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10.1016/0378-1119(92)90608-R

Cite this

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title = "Single-step purification of bacterially expressed polypeptides containing an oligo-histidine domain",

abstract = "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.",

keywords = "Affinity chromatography, Escherichia coli, expression vector, iminodiacetic acid-agarose, plasmid pUC19, recombinant DNA",

author = "{Van Dyke}, {Michael W.} and Mario Sirito and Mich{\`e}le Sawadogo",

note = "Funding Information: We are grateful to Alberto Fernandez, Paul Hardenbol, Kathy Heacock and Qun Lin for excellent technical assistance. This work was supported by grant No. 3110 from the Council for Tobacco Research (M.W.V.D.) and grant G-1195 from The Robert A. Welch Foundation (M.S.).",

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doi = "10.1016/0378-1119(92)90608-R",

language = "English (US)",

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pages = "99--104",

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T1 - Single-step purification of bacterially expressed polypeptides containing an oligo-histidine domain

AU - Van Dyke, Michael W.

AU - Sirito, Mario

AU - Sawadogo, Michèle

N1 - Funding Information: We are grateful to Alberto Fernandez, Paul Hardenbol, Kathy Heacock and Qun Lin for excellent technical assistance. This work was supported by grant No. 3110 from the Council for Tobacco Research (M.W.V.D.) and grant G-1195 from The Robert A. Welch Foundation (M.S.).

PY - 1992/2/1

Y1 - 1992/2/1

N2 - 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.

AB - 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.

KW - Affinity chromatography

KW - Escherichia coli

KW - expression vector

KW - iminodiacetic acid-agarose

KW - plasmid pUC19

KW - recombinant DNA

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AN - SCOPUS:0026603594

SN - 0378-1119

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EP - 104

JO - Gene

JF - Gene

IS - 1

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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