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@ARTICLE{Gupta:863044,
      author       = {Gupta, Saurabh and Sarkar, Sounik and Katranidis,
                      Alexandros and Bhattacharya, Jaydeep},
      title        = {{D}evelopment of a {C}ell-{F}ree {O}ptical {B}iosensor for
                      {D}etection of a {B}road {R}ange of {M}ercury {C}ontaminants
                      in {W}ater: {A} {P}lasmid {DNA}-{B}ased {A}pproach},
      journal      = {ACS omega},
      volume       = {4},
      number       = {5},
      issn         = {2470-1343},
      address      = {Washington, DC},
      publisher    = {ACS Publications},
      reportid     = {FZJ-2019-03166},
      pages        = {9480 - 9487},
      year         = {2019},
      abstract     = {Mercury (Hg) is one of the main water contaminants
                      worldwide. In this study, we have developed both whole-cell
                      and cell-free biosensors to detect Hg. Genetically modified
                      plasmids containing the merR gene were used to design
                      biosensors. Firefly luciferase (LucFF) and emerald green
                      fluorescent protein (EmGFP) genes were separately introduced
                      as a reporter. Both constructs showed a detection limit of 1
                      ppb (Hg) in Escherichia coli and the cell-free system. We
                      found that higher concentrations of Hg become detrimental to
                      bacteria. This cytotoxic effect shows an anomalous result in
                      high Hg concentrations. This was also observed in the
                      cell-free system. We found that EmGFP fluorescence was
                      decreased in the cell-free system because of a change in pH
                      and quenching effect by Hg excess. Once the pH was adjusted
                      to 7 and a chelating agent was used, the EmGFP fluorescence
                      was partially restored. These adjustments can only be done
                      in the cell-free system after the GFP expression and not in
                      whole cells where their number has been decreased because of
                      toxicity. Therefore, we suggest the use of the
                      cell-free-system, which not only reduces the total
                      experimental time but also allows us to perform these
                      postexperimental adjustments to achieve higher sensitivity.
                      We would also recommend to perform more measurements at a
                      time with different dilution factors to bring down the Hg
                      concentration within the measurable limits or to use some
                      other chelating agents which can further reduce the excess
                      Hg concentration.},
      cin          = {ICS-5},
      ddc          = {660},
      cid          = {I:(DE-Juel1)ICS-5-20110106},
      pnm          = {551 - Functional Macromolecules and Complexes (POF3-551)},
      pid          = {G:(DE-HGF)POF3-551},
      typ          = {PUB:(DE-HGF)16},
      UT           = {WOS:000473407200003},
      doi          = {10.1021/acsomega.9b00205},
      url          = {https://juser.fz-juelich.de/record/863044},
}