Please use this identifier to cite or link to this item:
Scopus Web of Science® Altmetric
Type: Journal article
Title: Purification of bacteriophage lambda repressor
Author: Gao, N.
Shearwin, K.
Mack, J.
Finzi, L.
Dunlap, D.
Citation: Protein Expression and Purification, 2013; 91(1):30-36
Publisher: Academic Press Inc Elsevier Science
Issue Date: 2013
ISSN: 1046-5928
Statement of
Ning Gao, Keith Shearwin, John Mack, Laura Finzi, David Dunlap
Abstract: Bacteriophage lambda repressor controls the lysogeny/lytic growth switch after infection of E. coli by lambda phage. In order to study in detail the looping of DNA mediated by the protein, tag-free repressor and a loss-of-cooperativity mutant were expressed in E.coli and purified by (1) ammonium sulfate fractionation, (2) anion-exchange chromatography and (3) heparin affinity chromatography. This method employs more recently developed and readily available chromatography resins to produce highly pure protein in good yield. In tethered particle motion looping assays and atomic force microscopy "footprinting" assays, both the wild-type protein and a C-terminal His-tagged variant, purified using immobilized metal affinity chromatography, bound specifically to high affinity sites to mediate loop formation. In contrast the G147D loss-of-cooperativity mutant bound specifically but did not secure loops. © 2013 Elsevier Inc. All rights reserved.
Keywords: Lambda repressor
Bacteriophage lambda
Heparin affinity
Tethered particle motion
Ammonium sulfate
Anion exchange
Single particle tracking
Rights: Copyright © 2013 Elsevier Inc. All rights reserved.
DOI: 10.1016/j.pep.2013.06.013
Published version:
Appears in Collections:Aurora harvest
Molecular and Biomedical Science publications

Files in This Item:
There are no files associated with this item.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.