- Why we choose Fec:
- + Conformational change
- + FecA, major component of the pathway, is an outermembrane protein
- + Fec pathway is non-essential
- + Well-understood pathway
- Our design:
- + System level: animation of how system is supposed to work
- + Device level: diagram with explanation
- Questions we address:
- Engineering/design of system:
- + internal fusion - image of FecA protein bound & unbound
- + signal transmission - flourescein --> PoPs --> expression of RFP
- + genes under polycistronic control
- + external linker added to anti-flourescein scFv
- Science of the system:
- + fusion positions - images of main 6, mention 17 total due to variation
- + knock out of native Fec system
- + device diagram of what parts are built and/or tested
- + results: data, graph, gel, microscope images
- + conclusions from present state of project:
- FecA Promoter-RFP is induced by iron
"What's left" is if project is not finished. If we've done building and tested system right away for its function and doesn't work, "Analysis of why system doesn't work."
- What's needed to be done:
- + parts to build/test
- + problems to solve
- + characterization of parts
- Analysis of why system doesn't work:
- + what could be the problems:
- - conformational change wasn't induced
- - no binding of flourescein
- - scFv wasn't expressed
Base on analysis above, "What's needed to be done" will be changed to tests that confirm/refute the causes above.
Questions for input
- other engineering questions?
- other scientific questions?
- data? transfer curve? FACS data? Building data vs. testing data
- under "what's left," characterization of parts overlapse with Jessica's direction.