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A GFP-based bacterial biosensor with chromosomally integrated sensing cassette for quantitative detection of Hg(II) in environment |
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| Authors: | Himanshu Priyadarshi Absar Alam Gireesh-Babu P Rekha Das Pankaj Kishore Shivendra Kumar and Aparna Chaudhari |
| Institution: | College of Fisheries (CAU), Lembucherra, Tripura 799210, India;Department of Fish Genetics and Biotechnology, Central Institute of Fisheries Education, Versova, Mumbai 400061, India;Department of Fish Genetics and Biotechnology, Central Institute of Fisheries Education, Versova, Mumbai 400061, India;Department of Fish Genetics and Biotechnology, Central Institute of Fisheries Education, Versova, Mumbai 400061, India;Department of Fish Genetics and Biotechnology, Central Institute of Fisheries Education, Versova, Mumbai 400061, India;Department of Fish Processing Technology, Central Institute of Fisheries Education, Versova, Mumbai 400061, India;Department of Fish Nutrition and Biochemistry, Central Institute of Fisheries Education, Versova, Mumbai 400061, India;Krishi Vigyan Kendra, Rajendra Agricultural University, Madhopur 845454, Bihar, India;Department of Fish Genetics and Biotechnology, Central Institute of Fisheries Education, Versova, Mumbai 400061, India |
| Abstract: | A mercury biosensor was constructed by integrating biosensor genetic elements into E. coli JM109 chromosome in a single copy number, using the attP/attB recombination mechanism of lambda phage. The genetic elements used include a regulatory protein gene (merR) along with operator/promoter (O/P) derived from the mercury resistance operon from pDU1358 plasmid of Serratia marcescens. The expression of reporter gene gfp is also controlled by merR/O/P. Integration of the construct into the chromosome was done to increase the stability and precision of the biosensor. This biosensor could detect Hg(II) ions in the concentration range of 100-1700 nmol/L, and manifest the result as the expression of GFP. The GFP expression was significantly different (P < or = 0.05) for each concentration of inducing Hg(II) ions in the detection range, which reduces the chances of misinterpretation of results. A model using regression method was also derived for the quantification of the concentration of Hg(II) in water samples. |
| Keywords: | attP/attB integration E coli biosensor GFP mercury |
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