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Team:LMU-Munich/Bacillus BioBricks
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==Bacillus BioBricks== | ==Bacillus BioBricks== | ||
Revision as of 12:25, 20 August 2012
The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".
[ more news ]
Bacillus BioBricks
We will create a toolbox of Bacillus BioBricks to contribute to the registry.
We would like to introduce Bacillus to the world of iGEM!!!
This Bacillus BioBrick Box contains compatible vectors and useful promoters.
Here is a list of all the vectors we cloned and used. Please note that pMAD is not in a BioBrick standard, but it is a useful tool to knock out genes inBacillus subtilis.
For the use of our vectors, please see our Protocols page. A general introduction to Bacillus subtilis and its integrative vectors can be found here. All vectors have ampicillin as E. coli resistance and RFP as selection marker.
| name | E. coli res. | B. subt. res. | insertion locus | description | derived from | reference |
| pSBBs1C-lacZ | Amp | Cam | amyE | with lacZ reporter gene | pAC6 | ? |
| pSBBs4S | Amp | Spec | thrC | empty | pDG1731 | ? |
| pSBBs1C | Amp | Cam | amyE | empty | pDG1662 | ? |
| pSBBs4S-PXyl | Amp | Spec | thrC | with Xylose-inducible promoter | pXT | ? |
| pSBBs3C-luxABCDE | Amp | Cam | sacA | with luxABCDE reporter cassette | pAH328 | ? |
| pSBBs0K-Pspac | Amp | Kan | replicative | with IPTG inducible promoter | pDG148 | ? |
| pSBBs2E | Amp | MLS | lacA | empty | pAX01 | ? |
The number in the vector's name codes for the insertion locus and the following letter for the Bacillus subtilis resistance gene according to the following table:
| number | insertion locus | letter | resistance |
| 0 | replicative | C | Chloramphenicol |
| 1 | amyE (amylase) | E | MLS (Erythromycin + Lincomycin) |
| 2 | lacA (laccase ?) | K | Kanamycin |
| 3 | sacA (saccase?) | S | Spectinomycin |
| 4 | thrC (threonine C) |
The concentrations of the antibiotics and the insertion tests can be found in our protocol section.
The promoters we submit:
Pveg, PliaI, PliaG, plepA, Pxyl, Pxyl+XylR
We also tested the Anderson promoter collection in Bacillus subtilis with pSBBs3C-luxABCDE. The data will be online in our data section soon.
Furthermore we plan to submit reporter genes optimized for Bacillus subtilis , namely GFP, lacZ, luc and mKate.
Useful for the registry are also 5 tages in Freiburg Standard (cMyc, 10His, Flag, Strep, HA).
More details to come soon.
