Construction of green fluorescent protein based bacterial biosensor for heavy metal remediation

Environmental contamination by heavy metals is a worldwide problem. Therefore, it is necessary to develop sensitive, effective and inexpensive methods, which can efficiently monitor and determine the level of hazardous metals in the environment. Conventional techniques to analyze metals, suffer from the disadvantages of high cost. Alternatively, development of simple system for monitoring heavy metals pollution is therefore needed. The present approach is based on the use of bacteria that are genetically engineered so that a measurable signal is produced when the bacteria are in contact with the bioavailable metal ions. Reporter genes are widely used as genetic tools for quantification and detection of specific cell population, gene expression and constructing whole cell biosensors as specific and sensitive devices for measuring biologically relevant concentrations of pollutants. An attempt has been made to construct the reporter gene enhanced green fluorescent protein and was expressed under the control of cadR gene, responsible for cadmium resistance. Recombinant strain Escherichia coli cadR30 was used, that carried cadR gene in pET30b expression vector and cloned. Clones confirmed by the expression of enhanced green fluorescent protein was detected under ultraviolet illumination and separated on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The construction of green fluorescent protein based Escherichia coli bacterial biosensor was developed based on green fluorescent protein expression under the control cadR gene of Pseudomonas aeruginosa BC15. The constructed bacterial biosensor is useful and applicable in determining the availability of heavy metals in soil and wastewater.