Difference between revisions of "Team:TUDelft/Demonstrate"
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<h1>A system for host-independent expression</h1> | <h1>A system for host-independent expression</h1> | ||
<br><p>Our team's mission was to set the basis for a universal mobile genetic tool which could be readily used to engineer a wide range of diverse bacterial species- contributing to a streamlined, host-independent multi-chassis Synthetic Biology. To achieve this, we set ourselves two main goals: the first goal was to investigate the implementation and viability of the phi29 bacteriophage linear DNA replication system as an orthogonal replication tool in bacterial cells. The second goal was to engineer a system which generates constant gene expression levels independently of transcriptional and transnational variations that are expected to occur when transferring circuits across organisms.</p> | <br><p>Our team's mission was to set the basis for a universal mobile genetic tool which could be readily used to engineer a wide range of diverse bacterial species- contributing to a streamlined, host-independent multi-chassis Synthetic Biology. To achieve this, we set ourselves two main goals: the first goal was to investigate the implementation and viability of the phi29 bacteriophage linear DNA replication system as an orthogonal replication tool in bacterial cells. The second goal was to engineer a system which generates constant gene expression levels independently of transcriptional and transnational variations that are expected to occur when transferring circuits across organisms.</p> | ||
| − | In this project, we have successfully demonstrated independence of expression levels from transcriptional variations with our novel incoherent feed-forward loop (IFFL) design, and have achieved cross-species expression level stabilization between <i>Escherichia coli</i> and <i>Pseudomonas putida</i>. Moreover, we have validated replication of our synthetic phi29 plasmid in cell-free systems as well as performed titration experiments to uncover the effects of different concentrations of phi29 replication-associated proteins in <i>E. coli</i> | + | <p>In this project, we have successfully demonstrated independence of expression levels from transcriptional variations with our novel incoherent feed-forward loop (IFFL) design, and have achieved cross-species expression level stabilization between <i>Escherichia coli</i> and <i>Pseudomonas putida</i>. Moreover, we have validated replication of our synthetic phi29 plasmid in cell-free systems as well as performed titration experiments to uncover the effects of different concentrations of phi29 replication-associated proteins in <i>E. coli</i></p> |
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Revision as of 10:03, 21 October 2019
A system for host-independent expression
Our team's mission was to set the basis for a universal mobile genetic tool which could be readily used to engineer a wide range of diverse bacterial species- contributing to a streamlined, host-independent multi-chassis Synthetic Biology. To achieve this, we set ourselves two main goals: the first goal was to investigate the implementation and viability of the phi29 bacteriophage linear DNA replication system as an orthogonal replication tool in bacterial cells. The second goal was to engineer a system which generates constant gene expression levels independently of transcriptional and transnational variations that are expected to occur when transferring circuits across organisms.
In this project, we have successfully demonstrated independence of expression levels from transcriptional variations with our novel incoherent feed-forward loop (IFFL) design, and have achieved cross-species expression level stabilization between Escherichia coli and Pseudomonas putida. Moreover, we have validated replication of our synthetic phi29 plasmid in cell-free systems as well as performed titration experiments to uncover the effects of different concentrations of phi29 replication-associated proteins in E. coli