Swimmy Bacteria : Chiba 2006

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Contents

Concept

Swimmy Bacteria -- the E.coli freeze and gather when they are caught by red light.
conceptSwimmy
red light on -> stop
red light off -> move
Control the E.coli's movement with light!

Design

to stop the e.coli by light, we needed a part that stop the e.coli, and combine this with a light receptor that UCSF made it last year.

there was some ways to stop the e.coli running:

  1. making a chemaera with chemotaxis receptor and light sensing receptor
  2. stop the motor protein's expression(motB protein: from last year's parts)
  3. controling chemotaxis

etc

we chose to control chemotaxis. We've found out, from the paper(..see refrences), the chemoreceptor that fix the e.coli tumble. it is a fragment of the chemoreceptor "Tsr" (a serin receptor)from aa 290 to 551. we call this tsr-cw(or tsr290).


pathway

Experiments

we made the new part rbs-tsrcw by PCR and made a new BioBrick (BBa_J29049).

Swarm Plate Assay

first we wanted to make sure this parts will work, so we put it suffix in the OmpC-promotor& araC-promotor. we found the OmpC promotor (R0082) was not working good, so then we used the araC promotor. -- results under construction --


Capillary Assay

we wanted to see if the tsr-cw is working good with this assay,too. we're waiting for the results...

Members

Chiba 2006 team あ

Experiments

light cannon

  • 10/15 we wrote a picture at the solid media.(not finished yet) also we changed the arabinose concentration at the liquid media.
  • 10/14 the Fluorescent lamp was the best light. we could see the on/off clearly in the solid media. we found that the thin media is better than the thicker one.
  • 10/11 we changed the arabinose & S-gal's concentration & the light sorce to find the best condition.

swimmy

  • 10/15 tried the capiraly assay to see chemotaxis. also we washed the e.coli and observed it with the microscope(but it was kind of hard)
  • 10/11 we're finding the suitable media to observe the E.coli's movement(chemotaxis).
  • 8/23 we made the tsr gene parts by PCR & discussed about the locked Tsr mutants.

Todo List

Referenses

Locked Tsr Mutants

  • A.Bren and M.Eisenbach (2000) -- How Signals Are Heard during Bacterial Chemotaxis: Protein-Protein Interactions in Sensory Signal Propagation (J.Bacteriol.)
  • P.Ames and J.S.Parkinson (1988) -- Transmembrane Signaling by Bacterial Chemoreceptors: E.coli Transducers with Locked Signal Output (Cell)
  • P.Ames and J.S.Parkinson (1994) -- Constitutively Signaling Fragments of Tsr, the Escherichia coli Serine Chemoreceptor (J.Bacteriol.)


Experiments

  • Microscope -- Real-time imaging of fluorescent flagellar filaments.J.bacteriol.2000
  • Chemotaxis Assay -- J.Adler -- A Method for Measuring Chemotaxis and Use of the Method to Determine Optimum Conditions for Chemotaxis by Escherichia coli
  • M.K.Slocum and J.S.Parkinson (1985) -- Genetics of Methyl-Accepting Chemotaxis Proteins in Escherichia coli: Null Phenotypes of the tar and tap Genes (J.Bacteriol.)
  • J.S.Parkinson (1976) --- cheA, cheB, and cheC of Escherichia coli and Their Role in Chemotaxis (J.Bacteriol.)

Last Update

  • 10/13 todo list
  • 10/11 swimmy & gene pathway
  • 8/23 refrences
  • 8/7 concept&members --maiko


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