IGEM:IMPERIAL/2008/Projects/B. subtilis Chassis

To B.subtilis or not to B.subtilis, that is THE question

[B.subtilis genome]

Vector

[pC194]

[pC194]

Given the activity of B.subtilis degrading E.coli derived plasmids, amybe it would be an idea to utilise PCR for plasmid production and selection of transformants in B.subtilis to by-pass this issue?

Promoters

Constitutive promoters:

Use of basal transcription promoters, potentially the promoter for one or various rRNAs, P3 promoter, RNAP subunit gene promoters and metablic gene promoters.

[P3 promoter]

[Annotated B.subtilis genome]

[Genome project]

[Metabolic Pathways of B.subtilis]

Probably ideal to pick a few (say 5) and characterise in order to find relative levels for use

  1 rRNA promoter
  1+ Metabolic related promoter (potential inducibility)
  1 RNAP subunit promoter
  P3 promoter
  Another basally transcribe sequence

All non-constiutive promoters should remain functional in B.subtilis though leaky (basal) transcription rate will however be different, the key promoter is the one at the start of the chain...

RBSs?

Needs research...