Team:LMU-Munich/Data
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[[File:Auswertung Anderson promoters.png|thumb|right|400px|Fig. 1: Luminescence measurement of Anderson promoters in the reporter vector pSB<sub>''Bs''</sub>3C-''luxABCDE''. OD<sub>''600''</sub> (right), LUMI (center) and OD<sub>''600''</sub> per LUMI (left) depending on the time (h) are shown for two different clones (green/blue). Data derive from three independent experiments. Curves were fitted over each other (t=0, OD<sub>''600''</sub>=0,3) and smoothed by taking average of three neighboring values.]] | [[File:Auswertung Anderson promoters.png|thumb|right|400px|Fig. 1: Luminescence measurement of Anderson promoters in the reporter vector pSB<sub>''Bs''</sub>3C-''luxABCDE''. OD<sub>''600''</sub> (right), LUMI (center) and OD<sub>''600''</sub> per LUMI (left) depending on the time (h) are shown for two different clones (green/blue). Data derive from three independent experiments. Curves were fitted over each other (t=0, OD<sub>''600''</sub>=0,3) and smoothed by taking average of three neighboring values.]] | ||
- | Eleven (J23100,J23101, J23102, J23103, J23106, J23107, J23113, J23114, J23115, J23117, J23118) of the nineteen promoters of the Anderson collection were evaluated in the reporter vector pSB<sub>''Bs''</sub>3C-''luxABCDE'' from the BioBrickBox containing the lux operon as a reporter for promoter activity. The gene expression which correlates to the promoter activity leads to the expression of the lux operon with the luciferase. The luminescence which is produced by the luciferase can be measured with the plate reader (BioTek). | + | Eleven (J23100,J23101, J23102, J23103, J23106, J23107, J23113, J23114, J23115, J23117, J23118) of the nineteen promoters of the [http://partsregistry.org/Part:BBa_J23100 Anderson collection] were evaluated in the reporter vector pSB<sub>''Bs''</sub>3C-''luxABCDE'' from the BioBrickBox containing the lux operon as a reporter for promoter activity. The gene expression which correlates to the promoter activity leads to the expression of the lux operon with the luciferase. The luminescence which is produced by the luciferase can be measured with the plate reader (BioTek). These promoters showed a very low acitivity in ''B. subtilis'' in comparison to the other ''Bacillus'' promoters. |
Revision as of 11:59, 17 September 2012
![Team-LMU Photo2.jpg](/wiki/images/a/a0/Team-LMU_Photo2.jpg)
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 ]
![Sporenfreunde](https://static.igem.org/mediawiki/2012/e/e3/Team_LMU_Sporenfreunde.png)
Data
Here you will find all of our project data. For our big breakthroughs, or to follow specific projects, see the individual project pages:
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![]() Bacillus BioBrickBOX |
![]() SporeCoat FusionProteins |
![]() Germination STOP |
Bacillus BioBrick Box - Promoters
![](/wiki/images/4/44/Auswertung_Anderson_promoters.png)
Eleven (J23100,J23101, J23102, J23103, J23106, J23107, J23113, J23114, J23115, J23117, J23118) of the nineteen promoters of the [http://partsregistry.org/Part:BBa_J23100 Anderson collection] were evaluated in the reporter vector pSBBs3C-luxABCDE from the BioBrickBox containing the lux operon as a reporter for promoter activity. The gene expression which correlates to the promoter activity leads to the expression of the lux operon with the luciferase. The luminescence which is produced by the luciferase can be measured with the plate reader (BioTek). These promoters showed a very low acitivity in B. subtilis in comparison to the other Bacillus promoters.
To measure the activity not only with the lux reporter operon, four promoters were cloned into the reporter vector pSBBs1C-lacZ to do beta-galactosidase assays and then to compare the results of the strength of these promoters in B. subtilis.
![](/wiki/images/e/e7/Auswertung_plate_reader_andere_promotoren.png)
The constitutive promoters PliaG and PlepA were evaluated in the reporter vector pSBBs3C-luxABCDE which contains the lux operon.
![](/wiki/images/b/b8/Englisch_Auswertung_PliaG_Pveg.png)
Germination Stop
We induced our germination-mutant strains to sporulate in Difco sporulation media. Then we measured the germination rate of mutant spores in a germination assay.
![](/wiki/images/8/8b/Germination_assay_bahhumbug.jpg)
Strains:
B40 -- cwlD::kan, sleB::mls, cwlJ::spec
B41 -- cwlD::kan, sleB::mls, gerD::cat
B42 -- cwlD::kan, cwlJ::spec, gerD::cat
B43 -- gerD::cat, sleB::mls, cwlJ::spec
B46 -- cwlD::kan, cwlJ::spec, gerD::cat, sleB::mls
B47 -- gerD::cat, sleb::mls, cwlJ::spec, cwlD::kan
The plate growth demonstrates the inability of our mutant spores to germinate. We can say that fewer than 1 out of 3x10^7 spores of strains B40, B41, B43, B46, and B47 germinated.