6th World Congress on Biotechnology
New Delhi, India
Banasthali University, India
Title: Real time PCR based expression analysis of metal resistance genes and quantification of Pseudomonas aeruginosa J007 strain in mine waste sample
Biography: Sangeeta Choudhary
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.