CPC C12Q 1/6897 (2013.01) [C12N 1/20 (2013.01); C12N 9/12 (2013.01); C12N 15/52 (2013.01); C12N 15/635 (2013.01); C12N 15/74 (2013.01); C12Y 207/13003 (2013.01); G21F 9/18 (2013.01); G21F 9/30 (2013.01); C12N 2830/002 (2013.01); C12N 2830/55 (2013.01); C12N 2840/002 (2013.01); C12N 2840/105 (2013.01); G01N 2520/00 (2013.01)] | 40 Claims |
1. A UO2F2 biosensor comprising a genetically engineered bacterial cell capable of heterologously and/or natively expressing histidine kinase 1363, and U-sensitive transcriptional regulator 1362;
wherein the bacterial cell is an engineered bacterial cell including a 1362 U-sensing genetic reportable molecular component and/or a 1362 U-sensing/U-neutralizing genetic molecular component, each comprising a U-sensitive promoter in a configuration wherein the U-sensitive promoter directly initiates expression of the U-sensing reportable molecular component and/or of the U-sensing U-neutralizing molecular component in presence of bioavailable U;
wherein the U-sensitive promoter is
a 1362 U-sensitive prompter comprising a U-sensitive transcriptional 1362 binding site having a DNA sequence
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in which
N1 is C or T;
N2 is G or A;
N3 is T or C;
N4 is C;
N5 is A or G;
N6 is G or C;
N7 C or G;
N8 is any nucleotide;
N9 is any nucleotide;
N10 is any nucleotide;
N11 is any nucleotide;
N12 is T or C;
N13 is G;
N14 is T or C;
N15 is C;
N16 is A or C;
N17 is G; and
N18 is C or G,
and in which N1 to N17 are selected independently
and wherein the genetically modified bacterial cell is an engineered bacterial cell further comprising an F-sensing riboswitch within at least one of
the U-sensing reportable genetic molecular component in a configuration wherein the U-sensing reportable genetic molecular component, is transcribed in presence of an effective amount of bioavailable fluoride,
an F-sensing reportable genetic molecular component in a configuration wherein the F-sensing reportable genetic molecular component is transcribed in presence of an effective amount of bioavailable fluoride, and
an F sensitive genetic circuit in which at least one molecular component is a reportable molecular component (and in particular one or more reportable genetic molecular component and/or one or more reportable cellular molecular component), the reportable molecular component expressed when the genetic circuit operates according to the circuit design in presence of bioavailable F.
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