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Sanger, F., Nicklen, S. and Coulson, A.R. (1977) DNA sequencing with chainterminating inhibitors, Proceedings of the National Academy of Sciences of the U.S.A., 74(12), 5463-5467.

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• TITLE: Cloning, expression, purification and characterization of replication protein from plasmid pGP2 from Acetobacter estunensis

  AUTHORS: Peter Grones, Jozef Grones

  KEYWORDS: Acetobacter Estunensis; Rep34 Protein; DNA-Binding Activity; ATPase Activity; Phosphatase Activity; Unwinding Activity

  JOURNAL NAME: Advances in Bioscience and Biotechnology, Vol.1 No.5, December 30, 2010

  ABSTRACT: The Acetobacter estunensis Rep34 protein participates in the replication of bacterial plasmid pGP2. The Rep34 protein of the A. estunensis, was cloned to the expression vector, that ensure fusion with a His-tag sequence (Rep34 His-tagged), over-expressed in Escherichia coli and purified by metal-affinity chromatography to yield a highly purified and active protein. On this purified protein number different activities and motifs were detected. DNA band-shift assays showed that the Rep34 His-tagged protein bound to the regulation region for replication on the linear double-stranded DNA. In the protein was determined phosphatase activity, ATPase activity and protein is possible to unwind double strand DNA.