"
Team:Imperial College London/Project/Switch/Overview
From 2011.igem.org
RebekkaBauer (Talk | contribs) |
|||
| Line 12: | Line 12: | ||
The third idea uses the same BioBrick parts from the 2008 Berkeley iGEM team to create a toxin/anti-toxin system to try and limit horizontal gene transfer. The Antiholin gene will be on the genome of the bacteria under the control of a strong promoter. The Holin and Lysozyme genes will be present on the same plasmid as the two auxin genes. The idea here is that the presence of the antiholin will prevent the cell from lysing from the effects of holin and lysozyme. In a different cell, i.e., one that does not have antiholin on its genome, the antiholin and lysozyme will kill the cell, preventing it from keeping the plasmid containing the auxin genes. | The third idea uses the same BioBrick parts from the 2008 Berkeley iGEM team to create a toxin/anti-toxin system to try and limit horizontal gene transfer. The Antiholin gene will be on the genome of the bacteria under the control of a strong promoter. The Holin and Lysozyme genes will be present on the same plasmid as the two auxin genes. The idea here is that the presence of the antiholin will prevent the cell from lysing from the effects of holin and lysozyme. In a different cell, i.e., one that does not have antiholin on its genome, the antiholin and lysozyme will kill the cell, preventing it from keeping the plasmid containing the auxin genes. | ||
| + | |||
| + | Cloning Strategy | ||
| + | |||
| + | The first step is to transform competent E.coli cells with the killswitch cassette from Berkeley 08, available in the registry. This will then give us many copies of the genes so that we can use them in transformations. | ||
| + | |||
| + | Next, we will need to obtain a source of the antiholin gene by itself, and this will be achieved using PCR. | ||
| + | |||
| + | The next, and quite possibly the hardest step will be to transform E.coli so that antiholin is present on the genome. For this, we will be generously helped by Rhys Algar. | ||
Revision as of 08:36, 4 August 2011
To help towards these issues, we have designed a few safety features that will go some of the way towards making this project safer.
Kill Switch Designs
The first idea was to implant a chemical into the selected region of soil that the bacteria would be unable to live without, but this was dismissed on environmental reasons as there is very little that we can add to the soil without damaging its composition or the organisms within.
The second idea was to implement the Holin/Anti-Holin regulated kill switch designed by the Berkeley 2008 iGEM team under the control of a UV sensitive promoter. Since UV light can only penetrate 0.3mm into soil, this would be an ideal method to ensure that the bacteria remain underground.
The third idea uses the same BioBrick parts from the 2008 Berkeley iGEM team to create a toxin/anti-toxin system to try and limit horizontal gene transfer. The Antiholin gene will be on the genome of the bacteria under the control of a strong promoter. The Holin and Lysozyme genes will be present on the same plasmid as the two auxin genes. The idea here is that the presence of the antiholin will prevent the cell from lysing from the effects of holin and lysozyme. In a different cell, i.e., one that does not have antiholin on its genome, the antiholin and lysozyme will kill the cell, preventing it from keeping the plasmid containing the auxin genes.
Cloning Strategy
The first step is to transform competent E.coli cells with the killswitch cassette from Berkeley 08, available in the registry. This will then give us many copies of the genes so that we can use them in transformations.
Next, we will need to obtain a source of the antiholin gene by itself, and this will be achieved using PCR.
The next, and quite possibly the hardest step will be to transform E.coli so that antiholin is present on the genome. For this, we will be generously helped by Rhys Algar.
