Difference between revisions of "Team:Birkbeck/Modeling"
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<h3>Modelling the directed evolution of bacteriophage lambda</h3> | <h3>Modelling the directed evolution of bacteriophage lambda</h3> | ||
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<p>To accomplish the goal of altering the host specificity of bacteriophage lambda, we took a step-by-step directed evolution approach, working according to the 'design - build - model - test - repeat' principle. This started with the design of several mutations both to alter the phage phenotype and to make segments of its genotype BioBrick-compatible. We then proceeded to 'build' individual genes through synthesis and cloning. The next steps are to recombine these genes into a recombinant bacteriophage genome, before carrying out testing and selection, and repeating this process.</p> | <p>To accomplish the goal of altering the host specificity of bacteriophage lambda, we took a step-by-step directed evolution approach, working according to the 'design - build - model - test - repeat' principle. This started with the design of several mutations both to alter the phage phenotype and to make segments of its genotype BioBrick-compatible. We then proceeded to 'build' individual genes through synthesis and cloning. The next steps are to recombine these genes into a recombinant bacteriophage genome, before carrying out testing and selection, and repeating this process.</p> | ||
Revision as of 10:34, 18 September 2015
Modelling
Modelling the directed evolution of bacteriophage lambda
To accomplish the goal of altering the host specificity of bacteriophage lambda, we took a step-by-step directed evolution approach, working according to the 'design - build - model - test - repeat' principle. This started with the design of several mutations both to alter the phage phenotype and to make segments of its genotype BioBrick-compatible. We then proceeded to 'build' individual genes through synthesis and cloning. The next steps are to recombine these genes into a recombinant bacteriophage genome, before carrying out testing and selection, and repeating this process.
- 1.
Short tail fibre (stf) and reporter genes are cloned into a GT11 bacteriophage lambda vector, while regulatory circuits are created to control both the transfer between lytic/lysogenic cycle and the production of the tail fibre assembly (tfa) protein, in order to prevent toxicity to the host cell. The circularised GT11 vector and plasmids containing the regulatory circuits are transformed into E.coli host cells.