Team:BABS UNSW Australia/project/containment
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Introduction
Due to the nature of our project, having several measures in place to ensure containment and biosafety is very important. We strongly felt that we should not experiment with introducing the invasion-listeriolysin devices into bacterial hosts until we had developed well characterised and effective safety checks. We focused on two key control measures: the Cre-lox recombination system which would delete genes encoding invasive proteins after entry into a cell, and a toxin-antitoxin system to contain bacteria to specific inducing agar plates and prevent horizontal gene transfer.
The Cre-Lox System
The Cre-lox system is an efficient method for imparting deletions, insertions and translocations at specific sites on a genome. These sites are described by a Lox sequence which consists of a 13bp recognition region followed by an 8bp spacer and then closed by another 13bp recognition sequence. Cre recombinase will recognise two of these sites and effect a recombination event between them. The outcome is dependent on the sequence orientation. This can be seen in the diagram below.
Why?
Design
Biobricks
The Toxin-Antitoxin System
The Toxin-antitoxin system can classically be looked at as a poison and an antidote-that is one protein will kill the cell, but the presence of the other protein will counteract it.
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