(1) We realized the quantitative measurement in DmpR response intensity using Renilla Luciferase. Compared to GFP team Peking-2013 used, we can detect DmpR response intensity more accurately.
Figure 1. DmpR circuit. (A) Peking-2013 (B) Jilin-2017
(2) Renilla Luciferase enabled us to conduct the measurement in vivo, which provided an easier and faster method in detecting.
Team Jilin_China in 2017 did several mutagenesis in sensor domain of DmpR. To compare the difference between Peking-2013 DmpR and our mutated DmpR downstreaming of the same constitutive promoter, we chose phenol, 2-CP, 4-CP, 2,4-DCP and pyrocatechol as the inducer. Expression level of Renilla Luciferase downstream of dmp operon (P0 promoter) was used to indicate the response intensity towards different phenolics. Comparisons were based on ratio of different DmpR response and mock's toward phenol, 2-CP, 4-CP, 2,4-DCP and pyrocatechol respectively.
Figure 2. The device we constructed to do comparison between peking-2013 dmpR and our mutated DmpR
It turned out that both DmpR can response to phenol, 2-CP, pyrocatechol. However, compared to Peking-DmpR, our DmR showed better response to phenol and pyrocatechol.
Figure 3. DmpR response sensitivity toward different phenolics. Jilin-2017-DmpR showed better response to phenol and pyrocatechol. ★，p<0.05