Site-specific DNA-affinity chromatography of the lac repressor
- 1 January 1980
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 8 (16) , 3721-3728
- https://doi.org/10.1093/nar/8.16.3721
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.Keywords
This publication has 24 references indexed in Scilit:
- Theoretical aspects of DNA-protein interactions: Co-operative and non-co-operative binding of large ligands to a one-dimensional homogeneous latticePublished by Elsevier ,2004
- Lac repressor binding to poly [d(A-T)]. Conformational changesBiochemical and Biophysical Research Communications, 1974
- Plasmid ColE1 as a Molecular Vehicle for Cloning and Amplification of DNAProceedings of the National Academy of Sciences, 1974
- The Location of the Repressor Binding Sites in the lac OperonProceedings of the National Academy of Sciences, 1974
- lac Repressor-operator interaction: IX. The binding of lac repressor to operators containing Oc mutationsJournal of Molecular Biology, 1974
- lac represser binding to non-operator DNA: Detailed studies and a comparison of equilibrium and rate competition methodsJournal of Molecular Biology, 1972
- The lac represser-operator interactionJournal of Molecular Biology, 1970
- lac repressor-operator interaction: I. Equilibrium studiesJournal of Molecular Biology, 1970
- SUPERCOILED CIRCULAR DNA-PROTEIN COMPLEX IN Escherichia coli : PURIFICATION AND INDUCED CONVERSION TO AN OPEN CIRCULAR DNA FORMProceedings of the National Academy of Sciences, 1969
- A Deoxyribonucleic Acid Polymerase from Micrococcus luteus (Micrococcus lysodeikticus) Isolated on Deoxyribonucleic Acid-CelluloseJournal of Biological Chemistry, 1968