Tianjin/DIODE
From 2007.igem.org
DesignBrief Introduction of Bio-diode
The diode, one of the most significant parts in the electric integrate circuit and electronic equipment, functions as allowing the current get through when the diode is positive-biased, and cut off while it is negative-biased. When the negative voltage increases to a threshold, the diode tends to be negatively broken down. In this project, the molecular signal AHL is considered as the current of electric circuit.Three types of cells: Generator cells、Amplifier cells and Block cells are immobilized into small beads with a diameter of 0.5 millimeter, then different beads are settled into particular parts of our specially designed equipment sequentially and respectively.The diode is build up with the amplifier cells and the block cells.The generator cells are thought as the electrical source and there are detector cells,which is considered as the lamp ,to detect the AHL.
On the contrary, if the positions of amplifier cells and block cells in our equipment are reversed, the AHL produced by generator cell would be degraded by block cells at the very beginning resulting in no amplification of the signal. Thus, hardly AHL could be detected in the outflow culture, a condition called ‘negative-biased block’.
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ModelWe construct a mathematical model to simulate the features of our bio-diode mainly by Ordinary Differential Equations. With the help of this model,we shows the ideal performance of each immobilized cells ,especially the amplifier cell, under different cell density and flux conditions.We also examine the features of different cells under the conditions of positive-biased, negative-biased and break-down. Thus the data and conclution derived from the model are quite helpful for our further wet experiment.
2. Model result 2.1 Model of Generator Cells | |||
ExperimentThe function of detector cells has to be defined first to build an relationship between the concentration of AHL and the fluorescence intensity. The experimental results show that at a range of AHL concentrations the concentration of AHL is proporational to the fluorescence intensity, so within this extent, the fluorescence intensity could be regarded as the value of output signal which is convenient to detect and quantitively defined. Then we test the functions of other three types of cells in immobilized beads by using the detector cells, proving that the generator and block cells works perfectly but the amplified cells should be improved due to the self-induction of its Lux Promoter. Finally, the three types of cells are fulfilled into our particular designed equipment whose functions are also researched carefully, and in order to solve the problem of amplifier cells, we transfer this part into another plasmid whose duplication is controlled by IPTG. Although the datas are not as our in silico model, it is substantiated that every part of our design has functioned well and the main characters of diode including the 'positively passing through', 'negatively blocked', 'break down' are realized by our equipment. 1.Datas and Graphs of Detector cells
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