Overproduction and Characterization of UvrA Protein Encoded by Plasmid from Acetobacter estunensis GP2
Department of Molecular Biology, Comenius University, Bratislava
Plasmid pGP2 from Acetobacter estunensis GP2 encodes 1 296 bp uvrA gene for 432 aa reparation protein exinuclease A. Bioinformatic analysis of the protein confirmed 13 ?-helixes and 14 ?-structures, two domains of ABC transporters, one GTPase domain and a 21-amino-acid large trans-membrane domain. The protein has 9888 % identities with some bacterial genome-encoded proteins. uvrA gene amplified by PCR was cloned in pET28a- expression vector and purified by HPLC affinity chromato¬graphy. The purified protein (47.17 kDa) shows the ATPase activity ten times lower than the Rep34 protein (isoletectric point 6.8) from plasmid pGP2.
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