5, 5, 10, 15, 30, 45, 60 min, after which, 0 05 pmol 5′-end fluor

5, 5, 10, 15, 30, 45, 60 min, after which, 0.05 pmol 5′-end fluorescein-labelled oligonucleotide (dT)35 was added. The samples were then loaded onto 2% agarose gels without ethidium bromide Quisinostat in vivo and separated by electrophoresis in a TAE buffer as described for EMSA tests. The incubation periods for each temperature, where 50% of (dT)35 was bound, were noted. Protein sequence analysis The amino acid sequences of studied SSB proteins were analyzed using standard protein–protein BLAST and RPS-BLAST. Multiple sequence alignment was generated in ClustalX, using a PAM 500 scoring matrix. The results were prepared using the GeneDoc editor program (http://​www.​psc.​edu/​biomed/​genedoc).

Acknowledgements This work was supported by Polish National Science Centre Grant NO. N/NZ1/01562 to M.N. References 1. Greipel J, Urbanke C, Maass G: The single-stranded DNA binding protein of Escherichia coli . Physicochemical properties and biological functions. In Protein-Nucleic Acid Interaction. Edited by: Saenger W, Heinemann U. London: Macmillan; 1989:61–86. 2. Alani E, Tresher R, KU55933 nmr Griffith JD, Kolodner RD: Characterization of DNA-binding and strand-exchange stimulation properties of y-RPA, a yeast single-strand-DNA-binding protein. J Mol Biol 1992, 227:54–71.PubMedCrossRef 3. Lohman TM, Overman LB: Two binding modes in Escherichia coli single strand binding protein-single

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