- 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
Continue to Accomplishments and & Submitted BioBricks