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The Scientific World Journal
Volume 2014, Article ID 593546, 14 pages
http://dx.doi.org/10.1155/2014/593546
Research Article

Characterization of the algC Gene Expression Pattern in the Multidrug Resistant Acinetobacter baumannii AIIMS 7 and Correlation with Biofilm Development on Abiotic Surface

1Institute of Bioinformatics and Biotechnology, University of Pune, Pune 411 007, India
2Ispat General Hospital, SAIL, Rourkela 769 005, India
3Department of Biotechnology, Shivaji University, Kolhapur 416 004, India
4Structural Bioinformatics Unit, Department of Biochemistry, Shivaji University, Kolhapur 416 004, India
5Dr. Babasaheb Ambedkar Marathwada University, Aurangabad 431 001, India

Received 30 July 2014; Revised 10 November 2014; Accepted 10 November 2014; Published 3 December 2014

Academic Editor: Jose Correa Basurto

Copyright © 2014 Praveen K. Sahu 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.

Supplementary Material

We performed molecular dynamic simulation on the 3-D model of the Acinetobacter baumannii AIIMS 7 PMM/PGM enzyme, which revealed data on the specific chemical interactions between metal ion (Mg2+) and the PMM/PGM protein. Moreover, the water-mediated Hydrogen bonds (water bridges) and hydrophobic interactions (Lennard-Jones) were also identified, as mentioned in the Supplementary Table S1. Regarding the genetic characterization of the algC gene of the MDR strain A. baumannii AIIMS 7, amplification of the algC gene from genomic DNA was performed using specifically designed primers (118_F / 1897_R, Table 1) and documented after anlaysis by agarose gel electrophoresis as demonstrated in supplementary Figure S1. Similarly, to verify the transcription of the algC gene, total RNA was isolated and two internal regions of algC ORF (1360 bp and 463 bp) were amplified from the cDNA using primer pair 363F / CD1701R, and internal nested primer pair 1260 nest1F / CD1701R respectively (Table 1); as illustrated in the supplementary Figure S2. After sequencing the 1781 bp algC gene, the in silico tool (Neural Network Promoter Prediction program http://www.fruitfly.org/seq_tools/promoter.html) was used to predict the promoter region of the gene, results of which has been depicted in the supplementary Figure S3. Regulatory elements of the algC gene such as promoter, ribosomal binding site (RBS), transcription start site, the -10 and -35 regions of the promoter, translation start and end, could be identified and annotated. supplementary Figure S4 shows the CLUSTALW alignment of target PMM/PGM sequence (AEC46864) from A. baumannii AIIMS 7 with the template sequence (1K2Y) from P. aeruginosa PMM/PGM S108A mutant. This alignment was very crucial for building the 3-D model of the PMM/PGM protein from A. baumannii AIIMS 7 and subsequent Molecular Dynamics simulation studies. The alignment of these two protein sequences revealed the respective metal binding residues, sugar binding residues and phosphate binding residues as illustrated using colored highlights.

  1. Supplementary Materials