Team:LMU-Munich/Data/Constitutive
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".
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Constitutive Bacillus Promoters
The constitutive promoters PliaG and PlepA were evaluated in the reporter vector pSBBs3C-luxABCDE which contains the lux operon. This is why the promoter activity leads to gene expression and to the production of the protein luciferase. The luminescence of this protein can be measured with the plate reader Synergy2 (Biotek) (Fig.1). All clones show a normal growth behaviour. The activity of the promoters increases during transition from log to stationary phase. The second clone of the promoters PlepA and PliaG did not show any luminescence activity. In the beginning of the growth curve the activity of both promoters increases to their maximum. This maximum activity appears during the same growth phase. PliaG has an activity maximum of about 100.000 Lumi/OD600 during the transition from logarithmic to the stationary phase. PlepA shows a maximum of about 400.000 Lumi/OD600. Comparing these two constitutive promoters the activity of PlepA is about four times higher than the activity of PliaG. In the late stationary phase the activity completely disappears.
The two constitutive promoters PliaG and Pveg were evaluated with the reporter vector pSBBs3C-lacZ which contains the lacZ reporter gene. Promoter activity leads to the expression of the β-galactosidase which directly correlates to the promoter activity. The β-galactosidase assay of the constitutive Bacillus promoters Pveg and PliaG was repeated three times. Data show one representative result (Fig.2). In the beginning of the growth curve both promoters show only low activity. But then it increases to a maximum before it decreases to the begininng level after about seven hours (Data not shown). Summing up the course of activity of both promoters Pveg and PliaG is very similar based on the growth curve. The highest β-galactosidase activity and therefore the highest activity of the promoter Pveg with a maximum of 65 Miller units can be found during the transition from the logarithmic to the stationary phase. This is about five times higher than the acitivity of the promoter PliaG with a maximum activity of about 12 Miller Units.