Purified Stx and λ phage initiator O proteins bind specifically to two different origins of replication in vitro.
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.
Department of Molecular Biology, University of Gdansk, Wita Stwosza 59, 80-308, Gdańsk, Poland. Electronic address: katarzyna.kozlowska@biol.ug.edu.pl.
Katarzyna I
KI
Kozłowska
Department of Molecular Biology, University of Gdansk, Wita Stwosza 59, 80-308, Gdańsk, Poland.
Joanna
J
Tymecka-Mulik
Department of Molecular Biology, University of Gdansk, Wita Stwosza 59, 80-308, Gdańsk, Poland.
Grzegorz
G
Węgrzyn
D010582:Bacteriophage lambda / Q000737:chemistry*; D004279:DNA, Viral / Q000737:chemistry*; D011485:Protein Binding; D018741:Replication Origin; D014764:Viral Proteins / Q000737:chemistry / Q000302:isolation & purification
D004279:DNA, Viral; C027812:O protein, Bacteriophage lambda; D014764:Viral Proteins
DNA-Protein interactions; Initiator O protein; Protein overproduction; Protein purification; Stx-phage; λ phage