Team:BIT/MO2
Abstracts:
One part of our aim of modeling is to verify the effect of signal molecules on the promoter and to make the biological expression process clearer and more organized. What’s more, the main purpose is to verify the feasibility of the experiments of expression group and guide the progress of the experiment more smooth. However, the expression of the gene line is relatively long, if the theory is wrong, and the mistakes are found after the experiment, there will be a great harm on the experiment schedule, and moreover, the change will be very difficult. Therefore, the feasibility of modeling validation before expression experiments can provide a theoretical basis for expression groups and provide guidance for the success of their subsequent experiments. In addition, the detection of biomolecules is usually difficult to achieve, because the amount of biomolecules in the human body is too small, so we considered using the double fluorescence system to achieve the amplification by mathematical model and detect the amount of material on . Therefore, we use the support of literature , use the model to verify the linear relationship between the GFP and RFP ratio, to verify that the dual fluorescence system can be used for our project detection link, which play a guiding experimental team successfully completed the role of the experiment.
Objective:
One part of our aim of modeling is to verify the effect of signal molecules on the promoter and to make the biological expression process clearer and more organized. What’s more, the main purpose is to verify the feasibility of the experiments of expression group and guide the progress of the experiment more smooth. However, the expression of the gene line is relatively long, if the theory is wrong, and the mistakes are found after the experiment, there will be a great harm on the experiment schedule, and moreover, the change will be very difficult. Therefore, the feasibility of modeling validation before expression experiments can provide a theoretical basis for expression groups and provide guidance for the success of their subsequent experiments. In addition, the detection of biomolecules is usually difficult to achieve, because the amount of biomolecules in the human body is too small, so we considered using the double fluorescence system to achieve the amplification by mathematical model and detect the amount of material on . Therefore, we use the support of literature , use the model to verify the linear relationship between the GFP and RFP ratio, to verify that the dual fluorescence system can be used for our project detection link, which play a guiding experimental team successfully completed the role of the experiment.
Introduction:
In this model, we made the following assumptions: (1) having constructed lysine-deficient Escherichia coli and obtained accurate experimental data (2) the promoter sequence unchanged after the inhibition of protein binding to the promoter. (3) the terminator can completely terminate the promoter (4) When the inhibitory protein is absent, the promoter can stably express red fluorescent protein (RFP) (5) don’t consider the expression of the leakage of gene lines.
The first part:
1.The gene line which produce green fluorescent
BIT_Figure_The gene line which produce green fluorescent
First, according to the gene line above, we can get the expression, transcription and degradation of the following genes. Among them, k1 is the producing rate of mRNAlacI, k-1 is the degrading rate of mRNAlacI, k2 is the producing rate of proteinlacI , k-2 is the degrading rate of proteinlacI :
BIT_Figure_Model 9
The expression formula of promoter activity was constructed by Hill equation. β1 is the maximum starting ability of the promoter (generally set to 1), M1 is the amount of leakage of the promoter, SluxR is the amount of regulation factor, kd is the SluxR value when the model change trend is changed:
BIT_Figure_Model 10
Finally, according to the differential equation to express the expression, transcription and degradation process of the whole gene line:
BIT_Figure_Model 12
ModelAccording to our gene line, it can be concluded that the amount of proteinlacI can be approximated as the amount of proteinGFP (green fluorescence).
BIT_Figure_Model 13
Among them, according to the database in the official website of igem , we can know the value of some parameters: the maximum capacity of start-up promoterβ1 = 0.6; the amount of the leakage of promoter M1 = 0; n1 = 2. So we put these known parameters into the equation and get the value of kd at equal distances: 50,100,150,200, finally draw a rough image:
BIT_Figure_The relationship between regulation factor and gene expression rate
From these differential equations, combining the gene line of the whole expression group,we can get the model of green fluorescent protein expression and get the conclusion: the concentration of lysine from the upstream is higher , the rate of survival of lysine-deficient Escherichia coli is higher , the fluorescence intensity of GFP is greater . And this is also consistent with the original intention of the team members who design this line diagram. We proved the feasibility of this gene line.
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