Team:Washington/Future
From 2009.igem.org
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# Test additional proteins in the new custom display construct (GFP, OpdA, etc) | # Test additional proteins in the new custom display construct (GFP, OpdA, etc) | ||
# Test designed proteins from FoldIt | # Test designed proteins from FoldIt | ||
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Revision as of 05:16, 15 October 2009
- 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 arabanose inducible promoter for better control over secretion system activation
- Combine with target vector so entire secretion system is contained in one plasmid.
- Display System
- Test additional proteins in the new custom display construct (GFP, OpdA, etc)
- Test designed proteins from FoldIt