Revision as of 10:02, 17 October 2009

Module 3: Genome Deletion

Overview

Project overview.

Module 3 is the final module of the system. The E.ncapsulator has successfully completed its job of drug production and packaging in a protective coating. Now, it needs to be prepared to be converted into a safe pill carrying the drug of interest. This is done by inducing bacterial cell death and removing all the potentially harmful genetic material.

Reusable module for removal of genetic material

Removal of genetic material by the use of restriction enzymes is important to prevent DNA transfer to unintended targets, creating bacteria for instance with higher survivability or more toxicity. This module is a highly reusable module for any chassis system where the chassis is no longer required after a certain stage and there is a need to remove genetic material after genes are expressed.

The genetic circuit

Under the control of a thermoinducible promoter (K098995), when the temperature is raised to 42°C, the restriction enzymes DpnII (K200009) and TaqI (K200010) are produced. These restriction enzymes cut frequently in the genome, digesting the entire genetic material and inducing cell death. The Dam enzyme (K200001) is used in its native methylation system to prevent leaky levels of restriction enzymes from killing the cell prematurely. The restriction enzyme cleavage properties and Dam methylation protection form an interesting balance.

Project Tour

Module 3 Contents

Restriction Enzymes

DAM Methylation

Genetic Circuit

Wet Lab

Modelling

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-Under the control of a thermoinducible promoter (K098995), when the temperature is raised to 42°C, the restriction enzymes DpnII (K200009) and TaqI (K200010) are produced.    These restriction enzymes cut frequently in the genome, digesting the entire genetic material and inducing cell death.  The native Dam methylation system is used to prevent leaky levels of restriction enzymes from killing the cell prematurely. The restriction enzyme cleavage properties and Dam methylation protection form an interesting system for further study.  <br>
+Under the control of a thermoinducible promoter (K098995), when the temperature is raised to 42°C, the restriction enzymes DpnII (K200009) and TaqI (K200010) are produced.    These restriction enzymes cut frequently in the genome, digesting the entire genetic material and inducing cell death.  The Dam enzyme (K200001) is used in its native methylation system to prevent leaky levels of restriction enzymes from killing the cell prematurely. The restriction enzyme cleavage properties and Dam methylation protection form an interesting balance.  <br>
-These restriction enzymes have similar recognition sequences, allowing for an overlap in cleavage capabilities.  When both restriction enzymes are expressed, there is comprehensive cutting, but when one enzyme becomes mutated and dysfunctional, the other restriction enzyme also works well by itself.
-However, restriction enzymes are highly toxic to the cell, where even minute amounts can induce destruction of the genome.  Therefore,
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Team:Imperial College London/M3

From 2009.igem.org