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Enzyme Research
Volume 2013, Article ID 597028, 4 pages
http://dx.doi.org/10.1155/2013/597028
Research Article

Cloning, Expression, and Purification of Nucleoside Diphosphate Kinase from Acinetobacter baumannii

Department of Biophysics, All India Institute of Medical Sciences, New Delhi 110029, India

Received 31 December 2012; Accepted 28 March 2013

Academic Editor: Munishwar Nath Gupta

Copyright © 2013 Juhi Sikarwar et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Acinetobacter baumannii is a multidrug resistant pathogenic bacteria associated with hospital acquired infections. This bacterium possesses a variety of resistance mechanisms which makes it more difficult to control the bacterium with conventional drugs, and, so far no effective drug treatment is available against it. Nucleoside diphosphate kinase is an important enzyme, which maintains the total nucleotide triphosphate pool inside the cell by the transfer of γ-phosphate from NTPs to NDPs. The role of nucleoside diphosphate kinase (Ndk) has also been observed in pathogenesis in other organisms. However, intensive studies are needed to decipher its other putative roles in Acinetobacter baumannii. In the present study, we have successfully cloned the gene encoding Ndk and achieved overexpression in bacterial host BL-21 (DE3). The overexpressed protein is further purified by nickel-nitrilotriacetic acid (Ni-NTA) chromatography.