Team:University College London/BioBricks

From 2012.igem.org

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(Module 4: Buoyancy)
 
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= BioBricks =
= BioBricks =
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== BioBricks submitted to the Registry ==
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<groupparts>iGEM012 University_College_London</groupparts>
== Module 1: Plastic Detection ==
== Module 1: Plastic Detection ==
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'''Figure 1:''' The figure above shows our genetic circuit for the Detection Module. The Constitutive Promoter (BBa_J23119) drives constant expression of the NahR protein BBa_K228004 ('''modified''' - see below). NahR is a transcriptional activator, which is activated in response to salicylate molecules. Our proxy for plastic - Persistant Organic Pollutants - have a salicylate-like domain, and are therefore capable of activating NahR. Activated NahR binds the p(sal) promoter (Figure 2) thus activating the circuit for Module 2 (Aggregation).  
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/1/16/Ucl2Dete.png" alt="Detection" /></div></html>
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'''Figure 1:''' The figure above shows our genetic circuit for the Detection Module. The Constitutive Promoter (BBa_J23119) drives constant expression of the NahR protein BBa_K228004 ('''modified''' - see below). NahR is a transcriptional activator, which is activated in response to salicylate molecules. Our proxy for plastic - Organic Pollutants - have a salicylate-like domain, and are therefore capable of activating NahR. Activated NahR binds the pSal promoter (Figure 2) thus activating the circuit for Module 2 (Aggregation).
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'''Modified BBa_K228004:''' The BBa_K228004 BioBrick must be modified as it combines both the NahR protein and the p(sal) promoter. We wish to seperate these.
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'''Modified BBa_K228004:''' The BBa_K228004 BioBrick must be modified as it combines both the NahR protein and the pSal promoter. We wish to seperate these.
Please see our [[Team:University_College_London/Module_1|Detection Description Subpage]] for more details on this Module.
Please see our [[Team:University_College_London/Module_1|Detection Description Subpage]] for more details on this Module.
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== Module 2: Aggregation ==
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/2/26/Ucligem201222Detection_BioBrick.png" alt="Detection" /></div></html>
 
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== Module 2: Aggregation ==
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/2/2a/UclCurli.png" alt="Aggregation" /></div></html>
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'''Figure 2:''' The Figure above illustrates our circuit for Module 2 (Aggregation). The p(sal) Promoter BBa_K228004 (modified - see below) which triggers the Aggregation circuit is turned on when bound by activated NahR protein from Module 1. Once the p(sal) promoter is activated, it drives expression of the Curli Cluster of genes (BBa_K540000). The individuals genes of the cluster are shown radiating from BBa_K540000.
 
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'''Modified BBa_K228004:''' The BBa_K228004 BioBrick must be modified as it combines both the NahR protein and the p(sal) promoter. We wish to seperate these.
+
'''Figure 2:''' The Figure above illustrates our circuit for Module 2 (Aggregation). The pSal promoter BBa_K228004 (modified - see below) which triggers the Aggregation circuit is turned on when bound by activated NahR protein from Module 1. Once the pSal promoter is activated, it drives expression of the curli cluster of genes (BBa_K729003). The individuals genes of the cluster are shown radiating from BBa_K540000.  
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For more information on how this circuit is triggered, and the individual roles of the Curli genes, please visit our [[Team:University_College_London/Module_2|Aggregation Description Subpage]]
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'''Modified BBa_K228004:''' The BBa_K228004 BioBrick must be modified as it combines both the NahR protein and the pSal promoter. We wish to seperate these.
 +
'''BBa_K729003:''' This BioBrick is modified from BBa_K540000, which has a cobalt promoter.
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/4/45/Ucligem2012Aggregation_BioBrick.png" alt="Aggregation" /></div></html>
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For more information on how this circuit is triggered, and the individual roles of the Curli genes, please visit our [[Team:University_College_London/Module_2|Aggregation Description Subpage]]
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== Module: Plastic Degradation ==
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== Module 3: Plastic Degradation ==
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/6/63/Ucligem2012Degradation_BioBrick.png" alt="Degradation" /></div></html>
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/8/89/UclDeg.png" alt="Degradation" /></div></html>
'''Figure 3:''' The image above shows the circuit for Module 3 (Degradation). The Constitutive Promoter (BBa_J23119) drives constant expression of the Laccase BioBrick (BBa_K729002). The Laccase gene we intend to use originates from a particular bacterial strain, and is capable of Polyethylene Degradation.
'''Figure 3:''' The image above shows the circuit for Module 3 (Degradation). The Constitutive Promoter (BBa_J23119) drives constant expression of the Laccase BioBrick (BBa_K729002). The Laccase gene we intend to use originates from a particular bacterial strain, and is capable of Polyethylene Degradation.
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For more information on how this circuit functions, please visit our [[Team:University_College_London/Module_3|Degradation Description Subpage]]
For more information on how this circuit functions, please visit our [[Team:University_College_London/Module_3|Degradation Description Subpage]]
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== Module: Buoyancy ==
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== Module 4: Buoyancy ==
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'''BBa_K729000  Temperature regulated Gas Vesicle Polycistronic gene cluster composite'''
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This composite consists of the gas vesicle polycistronic gene regulated by heat shock promoter BBa_K338001, repressor tetR and corresponding promoter P(TetR), such that a reduction in temperature induces greater expression of the gvp gene cluster. Activation of Heat Shock Promoter, triggers expression of the repressor protein tetR, which binds and repressors the promoter p(TetR).  p(TetR) is the promoter driving constitutive gas vesicle formation, and so activation of the repressor system down regulates gas vesicle formation.
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/4/47/Ucligem2012Buoyancy_BioBrick.png" /></div></html>
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== Module: Salt Tolerance ==
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[[File:BOUYANCYGFP.png|center|900px]]
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'''BBa_K729001 irrE form Deinococcus radiodurans confers salinity and osmotic stress tolerance'''
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irrE confers salt tolerance in E. coli. Sequence pending
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/3/32/Ucligem2012Salt_Tolerance_BioBrick.png" alt="Salt Tolerance" /></div></html>
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[[File:BOUY.png|center|900px]]
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'''Figure 4:''' The image above depicts the circuit for Module 4 (Buoyancy). The Starvation Promoter (BBa_K118011) drives expression of theT7 RNA polymerase when the environmental glucose concentration is low, then T7 RNA polymerase produced binds the T7 promoter resulting in the expression of GFP. As a future vision we will include the Gas Vesicle Cluster instead of the reporter gene. The Gas Vesicle Cluster selected originates from ''Bacillus Megaterium'', and induces buoyancy in the cell.
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For more information on how this circuit functions, please visit our [[Team:University_College_London/Module_4|Buoyancy Description Subpage]]
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Gas Vesicle Cluster
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== Module 5: Salt Tolerance ==
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/d/d0/UclSalt.png" alt="Salt Tolerance" /></div></html>
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'''Figure 5:''' The image above illustrates the circuit design for Module 5 (Salt Tolerance). The Constitutive Promoter (BBa_J23119) will drive constant expression of our novel BioBrick IrrE, which confers salt resistance onto ''E. Coli''.
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For more information on how this circuit functions, please visit our [[Team:University_College_London/Module_5|Salt Tolerance Description Subpage]]
== Module: Containment ==
== Module: Containment ==
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'''BBa_K729010, BBa_K729009 Three fold active biological containment System'''
 
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/b/b0/Ucligem2012Containment_BioBrick.png" alt="Containment" /></div></html>
 
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<html><div align="center"><img src="http://2012.igem.org/wiki/images/f/f9/UclContainment.png" alt="Containment" /></div></html>
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'''Figure 6:''' The above image shows the genetic circuits for our threefold Containment module. Plasmid A carries the three toxin genes - Holin/Endolysin (BBa_K729009), Colicin E3 (BBa_K729005), and EcoRI (BBa_K729006). Plasmid B carries the antitoxins Anti-Holin (BBa_K729010), Colicin E3 Immuntiy (BBa_K729008), and Methytransferase Ecori. Plasmid A and B work in conjunction with one another to prevent Horizontal Gene Transfer by conjugation. Plasmid C carries a periplasmic nuclease (BBa_K279004) driven by the constitutive promoter BBa_J23119.
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For more information on how this circuit functions, please visit our [[Team:University_College_London/Module_6|Containment Description Subpage]]
{{:Team:University_College_London/templates/foot}}
{{:Team:University_College_London/templates/foot}}

Latest revision as of 03:07, 27 September 2012

Contents

BioBricks

BioBricks submitted to the Registry

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Module 1: Plastic Detection

Detection

Figure 1: The figure above shows our genetic circuit for the Detection Module. The Constitutive Promoter (BBa_J23119) drives constant expression of the NahR protein BBa_K228004 (modified - see below). NahR is a transcriptional activator, which is activated in response to salicylate molecules. Our proxy for plastic - Organic Pollutants - have a salicylate-like domain, and are therefore capable of activating NahR. Activated NahR binds the pSal promoter (Figure 2) thus activating the circuit for Module 2 (Aggregation).


Modified BBa_K228004: The BBa_K228004 BioBrick must be modified as it combines both the NahR protein and the pSal promoter. We wish to seperate these.


Please see our Detection Description Subpage for more details on this Module.

Module 2: Aggregation

Aggregation


Figure 2: The Figure above illustrates our circuit for Module 2 (Aggregation). The pSal promoter BBa_K228004 (modified - see below) which triggers the Aggregation circuit is turned on when bound by activated NahR protein from Module 1. Once the pSal promoter is activated, it drives expression of the curli cluster of genes (BBa_K729003). The individuals genes of the cluster are shown radiating from BBa_K540000.

Modified BBa_K228004: The BBa_K228004 BioBrick must be modified as it combines both the NahR protein and the pSal promoter. We wish to seperate these.

BBa_K729003: This BioBrick is modified from BBa_K540000, which has a cobalt promoter.

For more information on how this circuit is triggered, and the individual roles of the Curli genes, please visit our Aggregation Description Subpage

Module 3: Plastic Degradation

Degradation

Figure 3: The image above shows the circuit for Module 3 (Degradation). The Constitutive Promoter (BBa_J23119) drives constant expression of the Laccase BioBrick (BBa_K729002). The Laccase gene we intend to use originates from a particular bacterial strain, and is capable of Polyethylene Degradation.

For more information on how this circuit functions, please visit our Degradation Description Subpage

Module 4: Buoyancy

BOUYANCYGFP.png


BOUY.png

Figure 4: The image above depicts the circuit for Module 4 (Buoyancy). The Starvation Promoter (BBa_K118011) drives expression of theT7 RNA polymerase when the environmental glucose concentration is low, then T7 RNA polymerase produced binds the T7 promoter resulting in the expression of GFP. As a future vision we will include the Gas Vesicle Cluster instead of the reporter gene. The Gas Vesicle Cluster selected originates from Bacillus Megaterium, and induces buoyancy in the cell.

For more information on how this circuit functions, please visit our Buoyancy Description Subpage

Gas Vesicle Cluster

Module 5: Salt Tolerance

Salt Tolerance


Figure 5: The image above illustrates the circuit design for Module 5 (Salt Tolerance). The Constitutive Promoter (BBa_J23119) will drive constant expression of our novel BioBrick IrrE, which confers salt resistance onto E. Coli.

For more information on how this circuit functions, please visit our Salt Tolerance Description Subpage

Module: Containment

Containment


Figure 6: The above image shows the genetic circuits for our threefold Containment module. Plasmid A carries the three toxin genes - Holin/Endolysin (BBa_K729009), Colicin E3 (BBa_K729005), and EcoRI (BBa_K729006). Plasmid B carries the antitoxins Anti-Holin (BBa_K729010), Colicin E3 Immuntiy (BBa_K729008), and Methytransferase Ecori. Plasmid A and B work in conjunction with one another to prevent Horizontal Gene Transfer by conjugation. Plasmid C carries a periplasmic nuclease (BBa_K279004) driven by the constitutive promoter BBa_J23119.

For more information on how this circuit functions, please visit our Containment Description Subpage