Team:NWU-China/Demonstrate
Demonstrate
In our project, although we verified the feasibility of biosensors, there is still a long way to go. According to the current experimental results, although the fluorescence induction of E. coli in Phe / Tyr is strongly linear (Fig.1, A & B), it takes 24 hours for the bacteria to reach steady state (Fig.1 C & D), and the time is too long. This means that biological test strips are not suitable for this, mainly because the bacteria grow slowly in the M9 medium, and urea and uric acid affect the growth of the bacteria (Fig.1 E & F).
Fig.2 (A) & (B) the 24th hour Phe / Tyrfluorescence induction
(C) & (D) Fluorescence induction curve of different concentrations of Phe/Tyr with time
(E) & (F) Effects of urea and uric acid on bacterial growth
The successful establishment of the colorimetric card shows the possibility that our projects could become more practical for the patient's home. We will replace GFP and RFP with fwYellow and amilCP (Fig.2) in the future, so that they do not need specific instruments (such as UV lamps and at home). Spectrophotometer, etc.) can know the approximate content of Phe and Tyr in the body. However, the biggest problem in our project is that the fluorescence intensity difference between GFP and RFP is too large. If we transfer the plasmid containing PtyrP-fwYellow and ParoF-amilCP in the same bacteria, the bacterial liquid will reach the steady state, and the bacterial liquid will be The color will only be yellow. Therefore, in the later experiments, we mutated and screened the PtyrP and ParoF promoters, and selected promoters with high expression and significant interaction with TyrR protein. We will also optimize media conditions and home shakers to make our systems easier and more accurate to use at home.
Fig.2 Phe/Tyr regulatory pathway