Evaluation of smtA expression and E. coli survival against cadmium ions

Abstract

Cadmium and other heavy metals lead to environmental danger, and these heavy metals are a great threat to human and other animal’s health. Investigation of the relationship between survival of E. coli and metallothionein smtA gene expression against cadmium ion is the goal of this research. Survival of recombinant bacteria containing smtA gene was analyzed against various concentrations of cadmium chloride salt using optical density (OD). At the resistive range, recombinant bacteria were subjected to different treatments. At the logarithmic phase of bacterial growth, sampling, RNA extraction and cDNA synthesis were performed and smtA gene expression was then analyzed by real-time PCR using designed primers for smtA gene and Amp resistance (as the calibrator gene). Relative gene expression was calculated using the ∆∆Ct method. The resistive range against cadmium chloride was 0.5–0.7 mM (minimum inhibitory concentration (MIC = 0.5 mM)). Survival and gene expression analysis showed that in induced bacteria, smtA expression was increased significantly that in turn conferred resistance to cadmium chloride prominently. There was a direct relationship between increased smtA gene expression and survival of the recombinant bacteria. Therefore, our result may help to confront to cadmium metal environmental pollution using overexpression of smtA gene expression in recombinant bacteria.