Construction of an extended range whole-cell tetracycline biosensor by use of the tet(M) resistance gene

Abstract

An extended range whole-cell tetracycline biosensor strain was constructed by insertion of the tet(M) gene, encoding tetracycline resistance by ribosomal protection, into plasmid pTGFP2, which contains a transcriptional fusion between a tetracycline regulated promoter and the green fluorescent protein gene. Tetracycline, oxytetracycline, chlortetracycline and minocycline all effectively induced the resulting Escherichia coli MC4100/pTGM biosensor and similar dose-response characteristics were recorded by flow cytometry for all four compounds. The novel tetracycline biosensor was responsive to drug concentrations ranging from below 5 ngml(-1) to 16 microgml(-1), which represents a significant improvement of the original version.