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@ARTICLE{Rupprecht:835978,
      author       = {Rupprecht, Christian and Wingen, Marcus and Potzkei, Janko
                      and Gensch, Thomas and Jaeger, Karl-Erich and Drepper,
                      Thomas},
      title        = {{A} novel {F}b{FP}-based biosensor toolbox for sensitive in
                      vivo determination of intracellular p{H}},
      journal      = {Journal of biotechnology},
      volume       = {258},
      issn         = {0168-1656},
      address      = {Amsterdam [u.a.]},
      publisher    = {Elsevier Science},
      reportid     = {FZJ-2017-05103},
      pages        = {25-32},
      year         = {2017},
      abstract     = {The intracellular pH is an important modulator of various
                      bio(techno)logical processes such as enzymatic conversion of
                      metabolites or transport across the cell membrane. Changes
                      of intracellular pH due to altered proton distribution can
                      thus cause dysfunction of cellular processes. Consequently,
                      accurate monitoring of intracellular pH allows elucidating
                      the pH-dependency of (patho)physiological and
                      biotechnological processes. In this context, genetically
                      encoded biosensors represent a powerful tool to determine
                      intracellular pH values non-invasively and with high
                      spatiotemporal resolution.We have constructed a toolbox of
                      novel genetically encoded FRET-based pH biosensors (named
                      Fluorescence Biosensors for pH or FluBpH) that utilizes the
                      FMN-binding fluorescent protein EcFbFP as donor domain. In
                      contrast to many fluorescent proteins of the GFP family,
                      EcFbFP exhibits a remarkable tolerance towards acidic pH
                      (pKa ∼ 3.2). To cover the broad range of physiologically
                      relevant pH values, three EYFP variants exhibiting pKa
                      values of 5.7, 6.1 and 7.5 were used as pH-sensing FRET
                      acceptor domains. The resulting biosensors FluBpH 5.7,
                      FluBpH 6.1 and FluBpH 7.5 were calibrated in vitro and in
                      vivo to accurately evaluate their pH indicator properties.
                      To demonstrate the in vivo applicability of FluBpH, changes
                      of intracellular pH were ratiometrically measured in E. coli
                      cells during acid stress.},
      cin          = {ICS-4 / IMET},
      ddc          = {540},
      cid          = {I:(DE-Juel1)ICS-4-20110106 / I:(DE-Juel1)IMET-20090612},
      pnm          = {552 - Engineering Cell Function (POF3-552) / 581 -
                      Biotechnology (POF3-581)},
      pid          = {G:(DE-HGF)POF3-552 / G:(DE-HGF)POF3-581},
      typ          = {PUB:(DE-HGF)25},
      pubmed       = {pmid:28501596},
      doi          = {10.1016/j.jbiotec.2017.05.006},
      url          = {https://juser.fz-juelich.de/record/835978},
}