Latest revision as of 03:16, 22 October 2009

Meetings Calendar

Metal Intake/Efflux

Introduction

The first step in our project is the uptake of the heavy metal cadmium (and cadmium only) into our B.subtilis cell. Therefore it would be logical to find a way in which we can increase the intake of cadmium without increasing the intake of other metals too.

It is known that a Bacillus subtilis cell takes up cadmium naturally through the manganese transport system. The manganese ion channel that we intend to either upregulate or control is the MntH ion channel. MntH is part of the Nramp family of proton-coupled metal ion transporters. MntH is regulated negatively by increasing manganese ion concentrations. This regulation ocurs via the promoter MntR. B.subtilis therefore has the ability to limit the manganese metal intake system when the intracellular concentration of Mn2+ starts to approach cytotoxic levels.

In B. Subtilis cation efflux is by an antiporter: the CadA efflux system. Cd, Co, Zn are pumped out by this ATPase dependant member of the CDF family.

We need to engineer our B.subtilis to be able to upregulate cadmium uptake and downregulate its efflux system in order to accumulate cadmium in the cell once the metal container decision has been made.

Modelling

Click to understand more about our Metal Intake Model

Equations

Click to view more of the Metal Intake Model equations

Results

Click to view the results of the Metal Intake Model

20 – 21 June 2009 - Europe workshop (London)
23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
23 October Practice Presentation (Newcastle)
23 October T-shirts are ready
27 October Practice Presentation (Sunderland)
27 October Poster is ready
30 October – 2 November 2009 - Jamboree (Boston)

Newcastle iGEM Twitter
[http://www.facebook.com/home.php#/group.php?gid=131709337641 Newcastle on Facebook]
[http://www.youtube.com/user/newcastle2009igem Newcastle Youtube Channel]

@@ Line 7: / Line 7: @@
 ==Introduction==
-The first step in our system would be to uptake the heavy metal cadmium ad cadmium only. Therefore it would be logical to find a way in which we can increase the intake of cadmium without increasing the intake of other metals too.
+The first step in our project is the uptake of the heavy metal cadmium (and cadmium only) into our ''B.subtilis'' cell. Therefore it would be logical to find a way in which we can increase the intake of cadmium without increasing the intake of other metals too.
-It is known that a Bacillus subtilis cell (from the 168 strain) takes up cadmium naturally through the manganese transport system (Laddaga. R.A., Bessen. R., Silver. S.; 1985). Additionally it has been demonstrated that mutations to the manganese transport system can affect its ability to uptake cadmium without affecting its ability to transport manganese (Zeigler. D.R., et al; 1987).
+It is known that a ''Bacillus subtilis'' cell takes up cadmium naturally through the manganese transport system.
+The manganese ion channel that we intend to either upregulate or control is the MntH ion channel. MntH is part of the Nramp family of proton-coupled metal ion transporters. MntH is regulated negatively by increasing manganese ion concentrations. This regulation ocurs via the promoter MntR. ''B.subtilis'' therefore has the ability to limit the manganese metal intake system when the intracellular concentration of Mn2+ starts to approach cytotoxic levels.
-The manganese ion channel that we intend to either upregulate or control is the mntH ion channel (also goes under the name ydaR). mntH is part of the Nramp family of proton-coupled, metal ion transporters (Que. Q., Helmann. J.D.; 2000). It is also classified as a secondary transporter (Membrane Transport website).
+In ''B. Subtilis''  cation efflux is by an antiporter: the CadA efflux system. Cd, Co, Zn are pumped out by this ATPase dependant member of the CDF family.
-It is also known that mntH is regulated negatively by increasing manganese ion concentrations. This is allowed to happen through the promoter called mntR (Que. Q., Helmann. J.D.; 2000). This means that B.subtilis has the ability to limit the manganese metal intake system when the intracellular concentrations of Mn+ starts to near cytotoxic levels.
+We need to engineer our ''B.subtilis'' to be able to upregulate cadmium uptake and downregulate its efflux system in order to accumulate cadmium in the cell once the metal container decision has been made.
-The cadA efflux system. In ''B. Subtilis''  cation efflux is by an antiporter. Cd, Co, Zn are pumped out by ATPase dependant systems of the CDF family.
 ==Modelling==
-* In normal time without any input from other systems, this is how the metal influx/efflux system works:<br>
+[[Image:Newcastle Metal intake1.png|center|400px]]
-<br>
-[[Image:Newcastle Metal intake1.png|center|500px]]
-<br>
+:''[[Team:Newcastle/Modelling/MetalIntake#Metal_Intake_Model| Click to understand more about our Metal Intake Model]]
-<br>
-* When the stochastic switch is giving feedback from it's metal container decision, this is how the system is engineered to work:
 <br>
-[[Image:Newcastle Metal intake2.png|center|500px]]
+===Equations===
-<br>
+[[Image:TeamNewcastleMetalIntakePicture1.png|400px|center]]
+:''[[Team:Newcastle/Modelling/MetalIntake#Equations| Click to view more of the Metal Intake Model equations]]
 <br>
-[[Image:Newcastle Metalintakesystem1.PNG|580px]]
+===Results===
+[[Image:Newcastle Metalintakemodel1.PNG|400px]]
+:''[[Team:Newcastle/Modelling/MetalIntake#Results| Click to view the results of the Metal Intake Model]]
+<br>
-*Here are some graphs which the produced by our cellML model
-[[Image:Newcastle Metalintakemodel1.PNG|580px]]
-[[Image:Newcastle Metalintakemodel2.PNG|580px]]
-[[Image:Newcastle Metalintakemodel3.PNG|580px]]
-==BioBrick constructs==
-[[Image:Newcastle Metal Intake Efflux Construct.png|center|500px]]
-==Lab Work Strategies==
-==Other Presentations and Diagrams==
 {{:Team:Newcastle/Footer}}
 {{:Team:Newcastle/Right}}

Team:Newcastle/Metalintakeefflux

From 2009.igem.org