US 12,134,795 B2
Sensor for cyanuric acid detection
Srivatsan Raman, Middleton, WI (US); and Xiangyang Liu, Madison, WI (US)
Assigned to Wisconsin Alumni Research Foundation, Madison, WI (US)
Filed by WISCONSIN ALUMNI RESEARCH FOUNDATION, Madison, WI (US)
Filed on Jan. 23, 2020, as Appl. No. 16/750,465.
Claims priority of provisional application 62/798,128, filed on Jan. 29, 2019.
Prior Publication US 2020/0239927 A1, Jul. 30, 2020
Int. Cl. C12N 15/62 (2006.01); C12N 1/20 (2006.01); C12N 15/113 (2010.01); C12Q 1/02 (2006.01); G01N 33/02 (2006.01); G01N 33/18 (2006.01); C07D 251/32 (2006.01)
CPC C12Q 1/02 (2013.01) [C12N 1/20 (2013.01); C12N 15/113 (2013.01); G01N 33/02 (2013.01); G01N 33/1826 (2013.01); C07D 251/32 (2013.01)] 15 Claims
 
1. A cyanuric acid-responsive biosensor, comprising
an E. coli host strain or an E. coli cell-free system comprising a first and a second expression cassette,
the first expression cassette comprising, in operable communication, a constitutive promoter and a gene encoding a Pseudomonas sp. AtzR cyanuric acid-responsive transcription factor, and
the second expression cassette comprising, in operable communication, a promoter regulated by the AtzR cyanuric acid-responsive transcription factor and a gene encoding a reporter protein,
wherein the first expression cassette expresses the AtzR cyanuric acid-responsive transcription factor, and, in the presence of cyanuric acid, the AtzR cyanuric acid-responsive transcription factor drives expression of the reporter protein, wherein the promoter regulated by the AtzR cyanuric acid-responsive transcription factor is a modified atzR-DEF promoter from a Pseudomonas sp. comprising an E. coli-10 sequence, an E. coli-35 sequence or both, and a 3′ E. coli constitutive σ70-dependent promoter,
wherein the E. coli-10 sequence is TATAAT, the E. coli-35 sequence is TTGACA, and the 3′ E. coli constitutive σ70-dependent proomoter is SEO ID NO: 19.