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Team:Washington/Future
From 2009.igem.org
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# Transfer to a Chlor resistance, p15A origin vector as used in the original papers | # Transfer to a Chlor resistance, p15A origin vector as used in the original papers | ||
# Add original upstream DNA (50bp) before the native RBS to ensure proper function | # Add original upstream DNA (50bp) before the native RBS to ensure proper function | ||
| - | # Add an | + | # Add an arabinose inducible promoter for better control over secretion system activation |
# Combine with target vector so entire secretion system is contained in one plasmid. | # Combine with target vector so entire secretion system is contained in one plasmid. | ||
Latest revision as of 04:09, 20 October 2009
Overview
- Target Construct
- Attempt to add additional proteins into the vector and test for functionality
- Vary linker lengths
- Make a simpler version for trouble shooting the secretion system: [6x-His]-[NheI]-[prtB]
- Transfer to a vector with the same origin and resistance as described in the original secretion system (pBR322+Carb).
- Add a lacI into the expression and target vectors so repression is hard-coded into the vector and expression can be induced regardless of the cell line.
- Secretion System
- Transfer to a Chlor resistance, p15A origin vector as used in the original papers
- Add original upstream DNA (50bp) before the native RBS to ensure proper function
- Add an arabinose inducible promoter for better control over secretion system activation
- Combine with target vector so entire secretion system is contained in one plasmid.
- Display System
- Construct a new modular display system: CDS
- Use computational protein design to create a monomeric protein that binds tightly to biotin: FoldIt
