An MFS Transporter-Like ORF from MDRAcinetobacter baumanniiAIIMS 7 Is Associated with Adherence and Biofilm Formation on Biotic/Abiotic Surface
Author(s) -
Praveen K. Sahu,
Pavithra S. Iyer,
Madhumita B. Gaikwad,
Sheetal C. Talreja,
Karishma R. Pardesi,
Balu A. Chopade
Publication year - 2012
Publication title -
international journal of microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.696
H-Index - 40
eISSN - 1687-9198
pISSN - 1687-918X
DOI - 10.1155/2012/490647
Subject(s) - biofilm , acinetobacter baumannii , open reading frame , microbiology and biotechnology , biology , transporter , heterologous expression , chemistry , gene , biochemistry , genetics , bacteria , peptide sequence , recombinant dna , pseudomonas aeruginosa
A major facilitator superfamily (MFS) transporter-like open reading frame (ORF) of 453 bp was identified in a pathogenic strain Acinetobacter baumannii AIIMS 7, and its association with adherence and biofilm formation was investigated. Reverse transcription PCR (RT-PCR) showed differential expression in surface-attached biofilm cells than nonadherent cells. In vitro translation showed synthesis of a ~17 kDa protein, further confirmed by cloning and heterologous expression in E. coli DH5 α . Up to 2.1-, 3.1-, and 4.1- fold biofilm augmentation was observed on abiotic (polystyrene) and biotic ( S. cerevisiae /HeLa) surface, respectively. Scanning electron microscopy (SEM) and gfp -tagged fluorescence microscopy revealed increased adherence to abiotic (glass) and biotic ( S. cerevisiae ) surface. Extracellular DNA(eDNA) was found significantly during active growth; due to probable involvement of the protein in DNA export, strong sequence homology with MFS transporter proteins, and presence of transmembrane helices. In summary, our findings show that the putative MFS transporter-like ORF ( pmt ) is associated with adherence, biofilm formation, and probable eDNA release in A. baumannii AIIMS 7.
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