Difference between revisions of "Team:Tsinghua-A/design of characters"

(Created page with "{{Tsinghua-A}} <html> <head> <meta charset="utf-8"> <meta http-equiv="X-UA-Compatible" content="IE=edge"> <title>Discription</title> <meta name="viewport" content="w...")
 
Line 141: Line 141:
 
&nbsp;&nbsp;&nbsp;&nbsp;This whole part is designed to produce a secretory invertase. As our <i>E. coli</i> can use glucose but not sucrose as its carbon source, farmers fulfill the role of providing nutrients.<br>
 
&nbsp;&nbsp;&nbsp;&nbsp;This whole part is designed to produce a secretory invertase. As our <i>E. coli</i> can use glucose but not sucrose as its carbon source, farmers fulfill the role of providing nutrients.<br>
 
<br>
 
<br>
&nbsp;&nbsp;&nbsp;&nbsp;However, from our <a href="zjx">orthogonality test</a> and <a href="ssl">killing test</a>, we find that we can just find warrior II that can be killed by warriors from the other group but not itself. Warrior I cannot satisfy this Therefore, we <a href="">improved our gene circuit by choosing specific combinations of AHL, receptor and promoter.</a></div>
+
&nbsp;&nbsp;&nbsp;&nbsp;However, from our <a href="https://2017.igem.org/Team:Tsinghua-A/orthogonality_test">orthogonality test</a> and <a href="https://2017.igem.org/Team:Tsinghua-A/killing_test">killing test</a>, we find that we can just find warrior II that can be killed by warriors from the other group but not itself. Warrior I cannot satisfy this Therefore, we <a href="https://2017.igem.org/Team:Tsinghua-A/fluid_model/improved_gene_circuit">improved our gene circuit by choosing specific combinations of AHL, receptor and promoter.</a></div>
 
<div class="myTitle2" >II. Design 2.0</div>
 
<div class="myTitle2" >II. Design 2.0</div>
 
<div class="myPic1" ><img src="https://static.igem.org/mediawiki/2017/d/d2/Designofcharactersfig2.png" width="559.333" height="191.333"></div>
 
<div class="myPic1" ><img src="https://static.igem.org/mediawiki/2017/d/d2/Designofcharactersfig2.png" width="559.333" height="191.333"></div>
 
<div class="myPicDis" >Fig. 2 Improved gene circuit. The one on the top is warrior I while the one below is warrior II.</div>           
 
<div class="myPicDis" >Fig. 2 Improved gene circuit. The one on the top is warrior I while the one below is warrior II.</div>           
<div class="myPara" >&nbsp;&nbsp;&nbsp;&nbsp;In this circuit, warrior II can only be killed by C4HSL secreted by warrior I, but not 3OC6HSL secreted by itself. (This can be shown in <a href="zjx">orthogonality test </a>)<br>
+
<div class="myPara" >&nbsp;&nbsp;&nbsp;&nbsp;In this circuit, warrior II can only be killed by C4HSL secreted by warrior I, but not 3OC6HSL secreted by itself. (This can be shown in <a href="https://2017.igem.org/Team:Tsinghua-A/orthogonality_test">orthogonality test </a>)<br>
&nbsp;&nbsp;&nbsp;&nbsp;Without TetR, we can see that C4HSL can activate expression of LacI by binding to RhlR and LuxR from results of <a href="zjx">orthogonality test </a>. However, we have TetR here! C4HSL forms a complex with the protein RhlR inside warrior I. This complex activate expression of TetR, TetR can then inhibit the promoter Plux/tetR, thus counteracting the self-activating effect. In this way, the expression of LacI is not activated so warrior I does not kill itself. We modify the gene circuits of beggar I and farmer I in a similar way so that warrior I no longer kill them. It feasibility can be seen in <a href="">Improved gene cicuit</a>.</div>
+
&nbsp;&nbsp;&nbsp;&nbsp;Without TetR, we can see that C4HSL can activate expression of LacI by binding to RhlR and LuxR from results of <a href="https://2017.igem.org/Team:Tsinghua-A/orthogonality_test">orthogonality test </a>. However, we have TetR here! C4HSL forms a complex with the protein RhlR inside warrior I. This complex activate expression of TetR, TetR can then inhibit the promoter Plux/tetR, thus counteracting the self-activating effect. In this way, the expression of LacI is not activated so warrior I does not kill itself. We modify the gene circuits of beggar I and farmer I in a similar way so that warrior I no longer kill them. It feasibility can be seen in <a href="https://2017.igem.org/Team:Tsinghua-A/fluid_model/improved_gene_circuit">Improved gene cicuit</a>.</div>
 +
 
  
 
   
 
   

Revision as of 08:47, 29 October 2017

Discription
Design of Characters
I. Design 1.0
Fig. 1 Design 1.0 of gene circuit
1. How warriors kill enemies:
    AHL1 in warrior I excretes AHL1. AHL1 goes into its enemies and forms a complex with the protein AHL1R. This complex activates the promoter Pahl1, thus activating the expression of LacI. The expression of LacI inhibits the promoter Plac, thus inhibiting the expression of cmR. Because we culture our E. coli in medium containing chloramphenicol, warrior I kills its enemies. As AHL1 does not form a complex with protein AHL2R inside E. coli from warrior I’s group, thus we fulfill our desire that warrior I just kill the enemies but not bacteria from it own group. Warrior Il works in a similar way.
2.How farmers provide nutrients:
    RafD encodes invertase, which can hydrolyze sucrose into glucose and fructose. The signal peptide of HlyA ligated to RafD can help the invertase be secreted through the recognition of its signal sequence. This is our new part. Besides, combining with extrinsic HlyB, HlyD and intrinsic TolC from the nucleoid in E. coli together form the transporter which helps the invertase to be secreted.
    This whole part is designed to produce a secretory invertase. As our E. coli can use glucose but not sucrose as its carbon source, farmers fulfill the role of providing nutrients.

    However, from our orthogonality test and killing test, we find that we can just find warrior II that can be killed by warriors from the other group but not itself. Warrior I cannot satisfy this Therefore, we improved our gene circuit by choosing specific combinations of AHL, receptor and promoter.
II. Design 2.0
Fig. 2 Improved gene circuit. The one on the top is warrior I while the one below is warrior II.
    In this circuit, warrior II can only be killed by C4HSL secreted by warrior I, but not 3OC6HSL secreted by itself. (This can be shown in orthogonality test )
    Without TetR, we can see that C4HSL can activate expression of LacI by binding to RhlR and LuxR from results of orthogonality test . However, we have TetR here! C4HSL forms a complex with the protein RhlR inside warrior I. This complex activate expression of TetR, TetR can then inhibit the promoter Plux/tetR, thus counteracting the self-activating effect. In this way, the expression of LacI is not activated so warrior I does not kill itself. We modify the gene circuits of beggar I and farmer I in a similar way so that warrior I no longer kill them. It feasibility can be seen in Improved gene cicuit.