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• Metal Intake/Efflux
• Metal Sensing
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Metal Intake/Efflux

Introduction

The first step in our project would be the uptake of heavy metal cadmium and cadmium only in 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.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.

In B. Subtilis cation efflux is by an antiporter: the cadA efflux system. Cd, Co, Zn are pumped out by this ATPase dependant systems 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 have been made.

Modelling

• In normal time without any input from other systems, this is how the metal influx/efflux system works:
  

• When the stochastic switch is giving feedback from it's metal container decision, this is how the system is engineered to work:

• Here are some graphs which the produced by our cellML model

BioBrick constructs

Lab Work Strategies

Other Presentations and Diagrams