IGEM:IMPERIAL/2008/Bioprinter/Subteam 1

B. subtilis and Motility: Subteam 1

OK. Lots of wetlabs hopefully, but also plenty of trawling for papers... =P

To Do

Cross these off when you've done them (or comment next to them) but don't delete them!

Contact

• Newcastle
• Hawaii
• Cambridge (?)
• Angelika

Dry Work

• Obtain strains
• Find Promoters
• Check ribosome binding sites (RBS)
• Protocols: cambridge 07
  • Culturing
  • Characterisation
  • 2nd transformation
  • Heterologous expression
• Find information on vectors
• Strain specifics
  • 168
  • PY79
• Information on integration loci

Wet Work

• Make those buggers work correctly
• etc.

Alternative Motility Strategies

Vincent was kind enough to point out a paper detailing different methods (apart from flagellar-based propulsion) of movement in B. subtilis; it focusses mainly on "sliding" but mentions "flagella-dependent swimming and swarming, and flagella-independent, twitching, gliding, and sliding" as the known methods of movement. The paper is below.

• A defined medium to investigate sliding motility in a Bacillus subtilis flagella-less mutant

Seems by "sliding" they mean... being pushed outwards by the expansion of the colony. The bacteria facilitate this by secreting chemicals that lower the friction between themselves and the surface. This shouldn't interfere with our goal too much, as any bacteria outside the target zone should stop secreting (and start swimming) anyway.

Motility Assays

We'll need to run these to gain parameters for macro-modelling of the system. From the literature it seems there are two basic classes; plate-based assays involving characterisation of the over-night colony size and shape and cell-tracking based assays monitoring the activity of a single cell under a microscope (time-lapse microscopy).

Vincent, again, was kind enough to find some papers on motility assays for bacteria (the Methodology sections of the following papers are worth a look)...

1. Comparative Analysis of the Development of Swarming Communities of Bacillus subtilis 168 and a Natural Wild Type: Critical Effects of Surfactin and the Composition of the Medium
2. Swarming motility in undomesticated Bacillus subtilis
3. MotPS is the stator-force generator for motility of alkaliphilic Bacillus, and its homologue is a second functional Mot in Bacillus subtilis
4. Properties of Motility in Bacillus subtilis Powered by the H+- coupled MotAB Flagellar Stator, Na+-coupled MotPS or Hybrid Stators MotAS or MotPB

...and also some software tools for carrying them out:

1. CellTrack: an open-source software for cell tracking and motility analysis
2. Automated measurement of cell motility and proliferation
3. Cell tracking software page
4. Cell Motion Tracking Software (Manual tracking)