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.