IGEM:IMPERIAL/2008/Projects/B. subtilis Chassis: Difference between revisions
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To ''B.subtilis'' or not to ''B.subtilis'', that is THE question | To ''B.subtilis'' or not to ''B.subtilis'', that is THE question | ||
[http://www.nature.com/nature/journal/v390/n6657/full/390249a0.html| ''B.subtilis'' genome] | |||
==Vector== | ==Vector== | ||
[http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=216434&blobtype=pdf| pC194] | |||
[http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=557300| 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? | 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? | ||
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Use of basal transcription promoters, potentially the promoter for one or various rRNAs, P3 promoter, RNAP subunit gene promoters and metablic gene promoters. | Use of basal transcription promoters, potentially the promoter for one or various rRNAs, P3 promoter, RNAP subunit gene promoters and metablic gene promoters. | ||
[http://jb.asm.org/cgi/reprint/189/5/1727.pdf| P3 promoter] | |||
[http://www.ncbi.nlm.nih.gov/sites/entrez?dispmax=1&db=nucleotide&doptcmdl=graph&term=50812173&from=9300&to=20924| Annotated ''B.subtilis'' genome] | |||
[http://www.ncbi.nlm.nih.gov/sites/entrez?Db=genomeprj&Cmd=Retrieve&list_uids=76| Genome project] | |||
[http://www.genome.jp/kegg/atlas/metabolism/2/| Metabolic Pathways of ''B.subtilis''] | |||
Probably ideal to pick a few (say 5) and characterise in order to find relative levels for use | 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 basaly 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... | 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... | ||
Revision as of 17:12, 9 July 2008
To B.subtilis or not to B.subtilis, that is THE question
Vector
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.
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 basaly 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...
