Difference between revisions of "Team:Nanjing-China/Model"

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       <ul><li><a href="https://2018.igem.org/Team:Nanjing-China/Model">Model</a></li></ul></div>
 
       <ul><li><a href="https://2018.igem.org/Team:Nanjing-China/Model">Model</a></li></ul></div>
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      <ul><li><a href="#intro">Introduction</a></li>
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      <li><a href="#method">Method</a></li>
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      <li><a href="#document">Document</a></li></ul>
 
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     <p>This year our team created a mathematical  model to optimize the arrangement of the nif gene cluster. This model helped we  optimized our design and provided some new perspectives of our  nitrogen-fixation system in transcriptional level.<br />
 
     <p>This year our team created a mathematical  model to optimize the arrangement of the nif gene cluster. This model helped we  optimized our design and provided some new perspectives of our  nitrogen-fixation system in transcriptional level.<br />
 
We developed this model with two goals in  mind:<br />
 
We developed this model with two goals in  mind:<br />
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     <p><font size="-1">Table1  The result of qPCR </font></p>
 
     <p><font size="-1">Table1  The result of qPCR </font></p>
 
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     <p> Method:<br />
 
     <p> Method:<br />
 
       To start with, we put all genes into two  groups. One group is under the strong promoter while the other is under the  weak one. We introduced some parameters shown in table2. </p>
 
       To start with, we put all genes into two  groups. One group is under the strong promoter while the other is under the  weak one. We introduced some parameters shown in table2. </p>
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       <p>With this arrangement, the proportion of nifB: nifH: nifD: nifK: nifE: nifN: nifX: nifV = 15.44: 46.93: 71.88: 62.10: 16.44: 16.04: 16.0: 15.94, which is most close to the ideal proportion among all the solutions.</p>
 
       <p>With this arrangement, the proportion of nifB: nifH: nifD: nifK: nifE: nifN: nifX: nifV = 15.44: 46.93: 71.88: 62.10: 16.44: 16.04: 16.0: 15.94, which is most close to the ideal proportion among all the solutions.</p>
 
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     Here is the codes we taped and used.
 
     Here is the codes we taped and used.
 
     <object width="100%" height="600px" data="https://static.igem.org/mediawiki/2018/7/7d/T--Nanjing-China--model-code.pdf" type="application/pdf">   
 
     <object width="100%" height="600px" data="https://static.igem.org/mediawiki/2018/7/7d/T--Nanjing-China--model-code.pdf" type="application/pdf">   

Revision as of 13:04, 11 October 2018

Nanjing-China2018

This year our team created a mathematical model to optimize the arrangement of the nif gene cluster. This model helped we optimized our design and provided some new perspectives of our nitrogen-fixation system in transcriptional level.
We developed this model with two goals in mind:
1.We want to achieve the best stoichiometric proportion of each nif gene, which is nifB:nifH:nifD:nifK:nifE:nifN:nifX:nifV=1:3:4:4:1:1:1:1.
2.We want our system as simple as possible, that means minimizing the number of promoters and copy number of each nif gene.
We made the following assumptions:
1.There are two kinds of promoters, both of which can successfully launch the expression of every nitrogen fixation gene involved in our system.
2.One promoter is stronger (called H) while the other is relatively weak(called L). Under promoter H, each gene’s transcription level is double that of under promoter L.
3.The order of genes has little influence on their transcriptional level.
We conducted Real-time Quantitative PCR to detect the transcription level of nif gene cluster and the experimental data we received became an important reference for our modeling.

 

gene

Average value of Cq

Relative expression level

16S DNA

6.33

 

nifB

19.97

7.80E-05

nifH

17.37

4.74E-04

nifD

18.34

2.42E-04

nifK

20.77

4.48E-05

nifE

22.20

1.66E-05

nifN

22.24

1.62E-05

nifX

22.92

1.01E-05

nifV

21.25

3.22E-05

Table1 The result of qPCR

Method:
To start with, we put all genes into two groups. One group is under the strong promoter while the other is under the weak one. We introduced some parameters shown in table2.

Parameters/data

Meanings

weak[ ]

the expression level of each nif gene under the weak promoter

strong[ ]

the expression level of each nif gene under the strong promoter

expected[ ]

the ideal stoichiometric proportion

d

deviation between the expected expression level and the actual expression level

Table 2

Then we did some necessary preprocessing. Firstly, we presumed the smallest element in each array was 1 and normalized all the other data accordingly. In addition, to ensure there is at least one solution, we adjusted expected[] to make each element greater than or equal to the smallest expression level of the corresponding gene.
After that, we began the organization. In order to minimize the total number of genes, we arranged the strong promoter group first, and considered the weak group later. For each gene, we constantly added one copy of it to the strong promoter group, calculated the current deviation after each addition and compared the current deviation with the last one. If the deviation was decreasing ,we added one more copy and repeated the operation until the last deviation was smaller than the current one. In that way, we were able to determine the number of each gene with which the deviations were the smallest and completed the arrangement of the strong group. Similarly, we arranged the weak group and finally received the result.

Fig 1. A flow diagram describing the idea of our modeling process

According to this flow diagram, we programmed with Python and got the following results:

Fig 2. The best arrangement of nif genes according to our calculation

With this arrangement, the proportion of nifB: nifH: nifD: nifK: nifE: nifN: nifX: nifV = 15.44: 46.93: 71.88: 62.10: 16.44: 16.04: 16.0: 15.94, which is most close to the ideal proportion among all the solutions.

Here is the codes we taped and used. TXT downlaod:https://static.igem.org/mediawiki/2018/f/fe/T--Nanjing-China--model.txt