Team:HKUST-Rice/Description
Project
Overview
Potassium (K), phosphorus (P) and nitrogen (N) are three plant macronutrients, and deficiencies in any of these can lead to plant diseases. By creating a biological sensor that can quickly provide soil status to plant owners, we can prevent plant diseases due to the lack of nutrients. In view of this, our team constructed a biological sensor in E. coli, which can detect KPN levels in the surrounding environment and give responses in the form of colors. In addition, we characterized the effects of a dual output system, in contrast to a single output system, in order to anticipate the expression of multiple outputs in a single system.
Potassium Sensor |
Phosphate Sensor |
Nitrate Sensor |
KdpFABC transporter is a high affinity K+ uptake system (Siebers, A. & Altendorf, K., 1988). The promoter upstream of kdpFABC operon,
kdpFp, works under low K+ concentrations (Polarek, J. W., et al., 1992; Walderhaug, M. O., et al., 1992). The goal is to characterize
kdpFp and build a device which is able to sense different concentrations of K+ and express different levels of GFP accordingly. |
phoAp and phoBRp promoters (Hsieh, Y. J., & Wanner, B. L., 2010) are cross-regulated by phoB and phoR, and are
usually repressed under high phosphate concentrations. phoR behaves as an activator as well as an inactivator for phoB.
When phosphate is limited, phoR will phosphorylate phoB and the phosphorylated phoB will directly activate the
phoAp and phoBRp promoters. |
yeaRp promoter (Lin, et al., 2007) is normally cross-regulated by the Nar two-component regulatory system (T. Nohno,
et al. , 1989) and NsrR, a regulatory protein. When there is nitrate and nitrite, it will be converted into nitric oxide.
The nitric oxide will bind to NsrR and relieve the repression on the yeaRp promoter. As a result, any
genes that are downstream of the yeaRp promoter will be expressed. |
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Signal Coexpression
In order to characterize the output difference between a single expression system and coexpression
from one vector, we constructed several inducible systems that give off fluorescence output and compared the dose
response of the individual systems in the two scenarios.
As part of the expression platform, we also aimed to construct a
3-input AND logic gate using toehold switches (Green, A. A., et al., 2014) to integrate the input signals detected from the three
(K, P and N) promoters.
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