Purified Stx and λ phage initiator O proteins bind specifically to two different origins of replication in vitro

Katarzyna Kozłowska , Joanna Tymecka-Mulik , Grzegorz Węgrzyn

Abstract

The O protein is a crucial factor initiating the DNA replication of lambdoid bacteriophage. Efficient DNA replication of Shiga toxin-converting phage is necessary for effective production of Shiga toxin – main virulence factor of STEC strains. We developed an improved protocol for overproduction, bacterial cell lysis and purification of λO protein. With use of this method we have also isolated O proteins of Stx-phage P27 and 933W that were never purified before. Purified proteins were tested for their DNA binding activity and revealed a sequence specific interactions.
Author Katarzyna Kozłowska KBM
Katarzyna Kozłowska ,,
- Department of Molecular Biology
, Joanna Tymecka-Mulik KBM
Joanna Tymecka-Mulik ,,
- Department of Molecular Biology
, Grzegorz Węgrzyn KBM
Grzegorz Węgrzyn,,
- Department of Molecular Biology
Journal seriesProtein Expression and Purification, ISSN 1046-5928
Issue year2017
Vol131
Pages16-26
Publication size in sheets0.5
Keywords in Englishprotein overproduction, protein purification, initiator O protein, DNA-protein interactions, λ phage, Stx-phage
DOIDOI:10.1016/j.pep.2016.11.002
URL http://dx.doi.org/10.1016/j.pep.2016.11.002
Languageen angielski
Score (nominal)15
ScoreMinisterial score = 15.0, 20-12-2017, ArticleFromJournal
Ministerial score (2013-2016) = 15.0, 20-12-2017, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 1.351 (2) - 2016=1.473 (5)
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