Cloning, expression, purification and characterization of replication protein from plasmid pGP2 from Acetobacter estunensis
Peter Grones, Jozef Grones
DOI: 10.4236/abb.2010.15055   PDF   HTML     11,080 Downloads   26,651 Views   Citations


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.

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Grones, P. and Grones, J. (2010) Cloning, expression, purification and characterization of replication protein from plasmid pGP2 from Acetobacter estunensis. Advances in Bioscience and Biotechnology, 1, 417-425. doi: 10.4236/abb.2010.15055.

Conflicts of Interest

The authors declare no conflicts of interest.


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