Team:BIOTEC Dresden/Project v2
From 2009.igem.org
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- | === | + | === Temporal and spatial control of protein synthesis by in vitro |
+ | recombination inside picoliter reactors === | ||
- | + | Manufacturing functionalized proteins in vitro poses a challenge, as | |
- | + | it requires coordinated molecular assemblies and multi-step reactions. | |
- | + | In this project we aim to control, over time and space, the production | |
- | + | of proteins tagged with a silver-binding peptide for in situ silver | |
- | + | nanoparticle nucleation inside microdroplets generated by microfluidic | |
- | + | devices. Combining a transcription-translation system with protein | |
- | + | coding genes and a recombination logic inside microdroplets provides | |
- | + | spatial control. Moreover, in the microfluidic chamber we can pinpoint | |
- | + | the beginning of synthesis, and easily track and isolate the droplets. | |
- | + | Site-specific recombination generates a molecular timer for temporal | |
- | + | control of protein synthesis. Unlike transcriptional regulation, this | |
- | + | method gives true all-or-none induction due to covalent modification | |
- | + | of DNA by Flp recombinase. Determining the transfer curve of inter-FRT | |
- | + | site distance versus average recombination time allows the onset of | |
- | + | gene expression to be predicted. We then apply this Flp reporter | |
- | + | system as a powerful PoPS measurement device. | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
{{:Team:BIOTEC_Dresden/NewTemplateEnd}} | {{:Team:BIOTEC_Dresden/NewTemplateEnd}} |
Revision as of 19:15, 19 September 2009
=== Temporal and spatial control of protein synthesis by in vitro recombination inside picoliter reactors ===
Manufacturing functionalized proteins in vitro poses a challenge, as it requires coordinated molecular assemblies and multi-step reactions. In this project we aim to control, over time and space, the production of proteins tagged with a silver-binding peptide for in situ silver nanoparticle nucleation inside microdroplets generated by microfluidic devices. Combining a transcription-translation system with protein coding genes and a recombination logic inside microdroplets provides spatial control. Moreover, in the microfluidic chamber we can pinpoint the beginning of synthesis, and easily track and isolate the droplets. Site-specific recombination generates a molecular timer for temporal control of protein synthesis. Unlike transcriptional regulation, this method gives true all-or-none induction due to covalent modification of DNA by Flp recombinase. Determining the transfer curve of inter-FRT site distance versus average recombination time allows the onset of gene expression to be predicted. We then apply this Flp reporter system as a powerful PoPS measurement device.