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From 2011.igem.org

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	** Solvent tolerance		** Solvent tolerance
		+	* Combine phage display with BioBrick vectors. Create a new vector that attaches the product of a BioBrick to a phage capsule protein. As proof of concept, combine this with random mutagenesis of some protein that attaches to some ligand, and screen for phage that attach the ligand. These phage will differ in their DNA sequences, but multiple rounds of reinsertion (a.k.a. biopanning) into the vector will find a winner. This is a form of directed evolution. Or as simpler proof of concept, just get phage display working with GFP or somesuch. This project is quite ambitious yet has achievable subgoals.
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Revision as of 23:08, 13 June 2011

Insert random project ideas here, no matter how crazy. These are not (yet) ideas we're seriously considering, but just whatever came into our heads at some point... Members may also wish to read the project ideas from last year.

• Last year's winner used DNA as a scaffold to latch proteins onto, via DNA binding domains. Could two such domains be used to link two replicons together, for purposes of increasing recombination frequency?

• E. coli lacks a Type II secretion system. (This is one of the ways bacteria can export a protein.) Add one! Test by export of a protein with signal added.
  • Comment from CF: actually, E. coli does possess a type II secretion system but it is normally inactive. When activated, it leads to secretion of a chitinase. You could use our BRIDGE system to make the necessary genetic modifications to have this pathway switched on in normal growth.

• Create a juxtacrine signalling pathway for E. coli. Probably impossible due to lipopolysaccharide. (Juxtacrine signalling is signalling by physical contact.)
  • Comment from CF: there have been some bizarre recent papers about bacteria being connected together by 'nanotubes', as well as a growing body of literature about electrically conductive pili (nanowires), but I'm not sure either of these systems is well enough characterized yet to form the basis of a project.
  • Comment from LK: I think this idea is similar to neural networks - http://en.wikipedia.org/wiki/Neural_network - if that was possible to model using E. coli, it would be awesome.

• (Chris French's idea) A sensor based on fusion of antibody domain to a signal transducing domain.

• Use the BRIDGE protocol to edit the chromosome of E. coli to report when a plasmid has been successfully introduced, e.g. by DNA binding proteins that would recognise the plasmid (either known sequences or common plasmid features like ORI) and cause some sort of effect.

• Global Transcription Machinery Engineering (see e.g. [http://www.sciencedirect.com/science/article/pii/S1096717606001248 Alper et al, 2007]) - use error-prone PCR to generate mutated copies of the rpoD gene (which codes for sigma-70, the key initiator of bacterial transcription). Transform these into cells. Because these mutant genes will affect transcription rates of many genes, some interesting phenotypes may be seen. Screen for e.g. solvent tolerance, heat and cold tolerance, etc. Interesting ones can be sequenced and biobricked. This project has natural quantitative aspects (how well can cells with various rpoD genes cope in various conditions) and may potentially generate multiple biobricks of use to others in the future.
  • Comment from AC: Alas, this may be impossible due to a [http://www.wipo.int/patentscope/search/en/WO2009061429 patent] on the technique.
    • But Chris says it's probably OK anyway.

• Instead of the above, clone sigma factors from various species into E. coli and screen for useful phenotypes.

• From our first brainstorm:
  • Bio-batteries
  • Bio-etching - i.e. making the bacteria eat through silicone plates
  • Bio-tweeting
  • Bio-sensors for soil acidity
  • Volcanic ash detection
  • Solvent tolerance

• Combine phage display with BioBrick vectors. Create a new vector that attaches the product of a BioBrick to a phage capsule protein. As proof of concept, combine this with random mutagenesis of some protein that attaches to some ligand, and screen for phage that attach the ligand. These phage will differ in their DNA sequences, but multiple rounds of reinsertion (a.k.a. biopanning) into the vector will find a winner. This is a form of directed evolution. Or as simpler proof of concept, just get phage display working with GFP or somesuch. This project is quite ambitious yet has achievable subgoals.

This was the old Navbox for Edinburgh; now it's obsolete...