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PMID: 7001362 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Site-specific DNA-affinity chromatography of the lac repressor.

Nucleic acids research ·Vol. 8 ·No. 16 ·1980-08-25 ·Pages 3721-8

Herrick G

Abstract

To test the feasibility of site-specific DNA-affinity chromatography, E. coli lac repressor was bound to an operator-containing DNA column, and in parallel to a non-operator DNA column. Salt gradient elution shows: 1) elution from non-operator DNA was near 250mM KCl or NaCl; interpretation of this result suggests the usefulness of the procedure for studying salt-dependence of DNA-protein affinities; 2) elution from operator-containing DNA was delayed (average elution = 1000mM salt), demonstrating a feasibility of site-specific DNA-affinity chromatography, if one provides a sufficiently favorable ratio of specific to non-specific DNA binding sites; 3) repressor eluted from operator-containing DNA over a very broad salt range, which may represent chromatography-generated repressor heterogeneity.

MeSH Terms
Chromatography, Affinity DNA, Bacterial Escherichia coli/analysis Kinetics Plasmids Repressor Proteins/isolation & purification Transcription Factors/isolation & purification
Chemicals
DNA, Bacterial Repressor Proteins Transcription Factors
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Herrick G
References (24)
24 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/8.16.3721
Published
1980-08-25
Pages
3721-8
Language
English
Region
England
NLM ID
0411011
PMCID
PMC324186
Subset
IM
Grants
NIGMS NIH HHS · GM25203 · United States
External Links
PubMed source DOI: 10.1093/nar/8.16.3721 PMC Full Text View journal details
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