Team:Cambridge/Signalling/Vectors
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Revision as of 03:59, 30 October 2008
All these vectors were kindly provided by Dr. Dan Ziegler, Director of the Bacillus Genetic Stock Center at Ohio State University.
Contents |
Integration Vectors
These vectors integrate into the chromosome and do not have a replication origin in Bacillus. They either integrate cassettes that require double crossovers and two homologous regions, or whole-plasmid insertions that only require one crossover and one corresponding region of homology.
Ectopic Integration
The integration site of these is pre-determined
BGSC Accession | Vector Name | Features | Integration Locus | Vector Map | Vector Sequence | Possible Uses | Necessity (1-5 stars) | Checked on Gel |
---|---|---|---|---|---|---|---|---|
ECE112 | pDG1661 |
| amyE | U46196 |
| **** | Yes | |
ECE171 | pPyr-Kan |
| pyrD | AY464559 |
| **** | Yes | |
ECE172 | pGlt-Cm |
| gltA | AY464560 |
| *: can use PyrD & AmyE instead | No: wrong size | |
ECE176 | pBCJ164 |
| rspD | BGSC sequence |
| *: use of strong rpsD promoter, but hard to insert into | Yes | |
ECE153 | pSG1154 |
| amyE | not available |
| ***: inducible promoter, strong gfp, no sequence | yes | |
ECE162 | pSG1193 |
| amyE | not available |
| ***: inducible promoter, strong gfp, no sequence | inconclusive : only one band |
Insert Integration
The user inserts a homologous piece of the Bacillus chromosome and the vector integrates there.
BGSC Accession | Vector Name | Features | Vector Map | Vector Sequence | Checked on gel |
---|---|---|---|---|---|
ECE149 | pMUTIN-GFP+ |
| From Bayreuth | No: 3 bands on the gel | |
ECE150 | pMUTIN-CFP |
| From Bayreuth (sequence incomplete) | yes | |
ECE151 | pMUTIN-YFP |
| From Bayreuth | Yes | |
ECE147 | pMUTIN-cMyc |
| From Bayreuth | yes |
Shuttle Vectors
BGSC Accession | Vector Name | Features | Vector Map | Vector Sequence | Checked on gel |
---|---|---|---|---|---|
ECE165 | pAD123 |
| BGSC sequence | Yes | |
ECE166 | pAD43-25 |
| BGSC sequence | Yes | |
ECE188 | pHCMC02 |
| From Bayreuth | Not yet checked, DNA stocks | |
ECE189 | pHCMC04 |
| From Bayreuth | Not yet checked, DNA stocks | |
ECE190 | pHCMC05 |
| From Bayreuth | Yes |
Plasmid Gels
- We ran these plasmids on gels, using double and single digests, to check them