Construction, Characterization, and Biosensing Potential of an SOS Inducible mCherry Based Reporter Plasmid
Construction, Characterization, and Biosensing Potential of an SOS Inducible mCherry Based Reporter Plasmid
Shaista Bano1, 2* and Sarfraz A. Tunio2
ABSTRACT
Improved variants of mushroom coral derived monomeric red fluorescent protein have been considered as a reliable reporter protein for gene expression. One of these variants is the mCherry protein which is encoded by the mCherry gene. The aim of the present study was to use the SOS inducible expression of the mCherry gene for monitoring the unwanted presence of antibiotics in a given environment. For that purpose, the mCherry gene was amplified using PCR technique. Then the amplified DNA fragment was cloned in a previously described low copy plasmid that carried an SOS inducible promoter. This cloning resulted in the transcriptional fusion between the SOS inducible promoter and the mCherry gene. The induced expression of the mCherry gene from the resultant construct was analyzed in E. coli cells by using fluorescence microscopy. The biosensing characteristics of the red fluorescent cells were evaluated quantitatively by considering the red fluorescence as an early marker of DNA damage in the cells exposed to antibiotics such as ColE7, nalidixic acid and norfloxacin. Our data showed that the construct was capable of detecting the presence of DNA damaging antibiotics up to nanogram level in an aqueous environment. Our findings suggested that the construct may be useful in the development of future strategies for the controlling of dissemination of antibiotic resistance that occurs due to the contamination of environment with antibiotic residues.
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