Team:LMU-Munich/Data

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Revision as of 10:07, 7 September 2012

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Data

On this page, we will upload data to explain our project progress. More to come!!!

Or see the results pages of our projects to follow our progress:

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Bacillus Intro	Bacillus BioBrickBOX	SporeCoat FusionProteins	Germination STOP

Bacillus BioBrick Box - Promoters

Fig. 1: luminescence measurement of Anderson promoters in the reporter vector pSB_Bs3C-luxABCDE. OD₆₀₀ (right), LUMI (center) and OD₆₀₀ per LUMI (left) depending on the time (h)are shown for two different clones (green/blue). Data derive from three independant experiments. Curves were fitted over each other (t=0, OD₆₀₀=0,3) and smothed by taking average of three neighbouring 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_Bs3C-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).

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.

Fig. 2: luminescence measurement of the constitutive Bacillus promoters P_liaG and P_lepA in the reporter vector pSB_Bs3C-luxABCDE. OD₆₀₀ (right), LUMI (center) and LUMI per OD₆₀₀ (left) depending on the time (h) are shown for two different clones (green/blue). Data come from three independant experiments. Curves were fitted over each other (t=0, OD₆₀₀=0,3) and smothed by taking average of three neighbouring values.

The constitutive promoters P_liaG and P_lepA were evaluated in the reporter vector pSB_Bs3C-luxABCDE which contains the lux operon.

Fig. 3: β-galactosidase assay and growth curve P_liaG (black) and P_veg (grey) in the reporter vector pSB_Bs1C-lacZ. β-galactosidase activity (Miller Units)and growth curve are the average of two independant clones. Experiment shows representative data which was recieved in the same way from three independant experiments.

Fig. 4:

Fig. 4:

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.