Real Time PCR Based Expression Analysis Of Metal Resistance Genes And Quantification Of Pseudomonas Aeruginosa J007 Strain In Mine Waste Sample | 38970
Journal of Biotechnology & Biomaterials
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A multi metal resistant bacterium Pseudomonas aeruginosa strain J007 isolated from uranium mine sites of Jaduguda, India
was characterized to detect the presence and expression of metal resistance determinants. PCR based detection followed
by sequence identity and phylogenetic analysis revealed presence of specific metal resistance genes copA and czcA in this isolate.
Real-time PCR based expression studies of these genes indicated a general up-regulation of both the genes in response to Cu,
Cd or Zn while sodA gene encoding superoxide dismutase enzyme was up-regulated only at higher metal concentrations. High
levels of mRNA transcripts of copA and czcA genes in response to specific metals suggest that these resistance systems have
important role in conferring metal resistance to the strain. Employing real-time PCR, 16S rRNA and czcA genes were targeted
to determine the abundance of P. aeruginosa J007 like strains in mine waste sample. Based on copy numbers of 16S rRNA and
czcA gene, abundance of P. aeruginosa J007 like strains was determined in the range of 1.5±0.7×104 to 3.1±0.98×104 cells g-1
sample. Real-time PCR based method developed in this study for detection of metal resistance genes as well as enumeration
of this kind of bacterium in contaminated samples using both structural and functional biomarkers could be an efficient
monitoring tool for in situ bioremediation.
Sangeeta Choudhary is an Assistant Professor in Department of Bioscience and Biotechnology, Banasthali University, Rajasthan, India. She has completed her PhD in 2010 from Indian Institute of Technology Kharagpur, India. She has published several papers in reputed peer reviewed journals. Her research interests include microbial diversity and gene expression under stress condition, metagenomics and bioremediation.