Idea 6/11

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(Experiments Required)
(Experiments Required)
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Urease vs. phosphatase.
Urease vs. phosphatase.
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#Solutions of CaCl2 and either phosphate ion donor (eg/P2O5) or carbonate ion donor (sodium bicarbonate?). See what the quality of the product is and how long it takes to form. Can start at a low concentration of either substrate and work up to a higher one. Can do it both ways round to find a minimum concentration of both required to precipitate calcium compound.  
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#Proteus vulgaris produces urease and colonies of P. vulgaris lower the pH by producing ammonia. A test can be done with Proteus vulgaris to see if this causes calcium to precipitate from the medium with some different concentrations of calcium ions. Locate an E. coli phosphatase and do a similar experiment, to see which compound is the 'best' solid material.
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#Locate urease/phosphatase in E. coli that is suitable. (possibly Alkaline phosphatase) Tests for conditions that will work. See how bacteria react with the ideal concentrations worked out in experiment one, (particularly for urease). CaCl2 concentrations, phosphate donor concentrations in medium. (Just test with genes coupled with constitutive promoters?) Can also see what happens when the medium, and if the medium becomes depleted of calcium or phosphate ions. End - Find a suitable ratio of calcium to phosphate ions in medium.  
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#Locate urease/phosphatase in E. coli that is suitable. Find optimum CaCl2 concentrations, phosphate donor concentrations in medium. (Just test with genes coupled with constitutive promoters?) Can also see what happens when the medium, and if the medium becomes depleted of calcium or phosphate ions. End - Find a suitable ratio of calcium to phosphate ions in medium.  
#Urease increases pH, measure pH change caused by different densities of bacteria, find if the pH change produced by E. coli in a lawn is suitable (high enough) for causing calcite to precipitate.  
#Urease increases pH, measure pH change caused by different densities of bacteria, find if the pH change produced by E. coli in a lawn is suitable (high enough) for causing calcite to precipitate.  
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Revision as of 12:11, 4 July 2006

Contents

Calcite/Calcium Phosphate Project (Idea 6/11)

Aim

To create a 3D structure in a defined area using bacteria which can precipitate out a calcium compound.


Background

Possible Methods

Initial colony makes phosphatase or urease in response to stimulus from UV, or absence of UV signal. The enzyme catalyses the conversion of calcium and phosphate/carbonate ions in the medium into calcium phosphate or calcium carbonate. An amplifier can be incorporated if the resulting amounts of 'hard' material need to be increased.


Problems

If two colonies are used then colony two has to be able to get to colony one. Maybe colony one could be non-motile and colony two motile?

How will the Calcium compound be restricted to only the areas activated? It could diffuse around the area and lose the original pattern.


Parts Required

UV responsive sensor

glycerol-3-phosphate synthesis gene (periplasmic/outer membrane?)

glycerol-3-phosphatase/urease protein coding gene


Experiments Required

Experiments for Idea 2.

Is calcium phosphate or calcite better at precipitating?

Urease vs. phosphatase.

  1. Proteus vulgaris produces urease and colonies of P. vulgaris lower the pH by producing ammonia. A test can be done with Proteus vulgaris to see if this causes calcium to precipitate from the medium with some different concentrations of calcium ions. Locate an E. coli phosphatase and do a similar experiment, to see which compound is the 'best' solid material.
  2. Locate urease/phosphatase in E. coli that is suitable. Find optimum CaCl2 concentrations, phosphate donor concentrations in medium. (Just test with genes coupled with constitutive promoters?) Can also see what happens when the medium, and if the medium becomes depleted of calcium or phosphate ions. End - Find a suitable ratio of calcium to phosphate ions in medium.
  3. Urease increases pH, measure pH change caused by different densities of bacteria, find if the pH change produced by E. coli in a lawn is suitable (high enough) for causing calcite to precipitate.


Ideas

Personal tools
Past/present/future years