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Team:LMU-Munich/Data/Inducible
From 2012.igem.org
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| - | [[File:plate reader induzierbar promotor.png|thumb|right|400px|<p align="justify">'''Fig. 1: Luminescence measurements of the promoter P<sub>''liaI''</sub> after induction with different concentrations of bacitracin.'''</p> ]] | + | [[File:plate reader induzierbar promotor.png|thumb|right|400px|<p align="justify">'''Fig. 1: Luminescence measurements of the inducible ''B. subtilis'' promoter P<sub>''liaI''</sub> after induction with different concentrations of bacitracin.''' OD600 (up), luminescence (middle), and luminescence/OD600 (down) of the two clones (green/blue) depending on the time (h) after induction with bacitracin (0 μg/ml(left) to 100μg/ml(right). Data derive from three independent experiments. t=0 is the time of induction.(</p> ]] |
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The inducible promoter '''P<sub>''liaI''</sub>''' was evaluated in the reporter vector pSB<sub>Bs</sub>3C-<i>luxABCDE</i> which contains the ''lux'' operon [[File:Lux operon.png|100px]]. 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'''). </p> | The inducible promoter '''P<sub>''liaI''</sub>''' was evaluated in the reporter vector pSB<sub>Bs</sub>3C-<i>luxABCDE</i> which contains the ''lux'' operon [[File:Lux operon.png|100px]]. 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'''). </p> | ||
| - | [[File:Englisch Auswertung PliaI.png|thumb|right|400px|<p align="justify">'''Fig. 2: β-galactosidase assay of the | + | [[File:Englisch Auswertung PliaI.png|thumb|right|400px|<p align="justify">'''Fig. 2: β-galactosidase assay and growth curve (with/without induction of bacitracin) of strains carrying the promoter P<sub>''liaI''</sub> fused to ''lacZ'''''. β-galactosidase activity (Miller Units)and the growth curve values are the average of two independant clones with their standard deviation that were measured during the same experiment. Experiment shows representative data which was obtained in the same way from three independent experiments.</p>]] |
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<p align="justify"> | <p align="justify"> | ||
The inducible promoter P<sub>''liaI''</sub> was also evaluated with the reporter vector pSB<sub>''Bs''</sub>1C-''lacZ'' which contains the ''lacZ'' reporter gene [[File:LacZ.png|50px]] ('''Fig.2'''). Promoter activity leads to the expression of the β-galactosidase which directly correlates to the promoter activity. The β-galactosidase assay of the inducible ''Bacillus'' promoter P<sub>''liaI''</sub> was repeated three times. Data show one representative result. | The inducible promoter P<sub>''liaI''</sub> was also evaluated with the reporter vector pSB<sub>''Bs''</sub>1C-''lacZ'' which contains the ''lacZ'' reporter gene [[File:LacZ.png|50px]] ('''Fig.2'''). Promoter activity leads to the expression of the β-galactosidase which directly correlates to the promoter activity. The β-galactosidase assay of the inducible ''Bacillus'' promoter P<sub>''liaI''</sub> was repeated three times. Data show one representative result. | ||
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| + | 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 P<sub>''veg''</sub> and P<sub>''liaG''</sub> is very similar based on the growth curve. The highest β-galactosidase activity and therefore the highest activity of the promoter P<sub>''veg''</sub> 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 P<sub>''liaG''</sub> with a maximum activity of about 12 Miller Units. | ||
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Revision as of 08:55, 24 September 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 ]
Inducible Bacillus Promoters
Fig. 1: Luminescence measurements of the inducible B. subtilis promoter PliaI after induction with different concentrations of bacitracin. OD600 (up), luminescence (middle), and luminescence/OD600 (down) of the two clones (green/blue) depending on the time (h) after induction with bacitracin (0 μg/ml(left) to 100μg/ml(right). Data derive from three independent experiments. t=0 is the time of induction.(
The inducible promoter PliaI was 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).
Fig. 2: β-galactosidase assay and growth curve (with/without induction of bacitracin) of strains carrying the promoter PliaI fused to lacZ. β-galactosidase activity (Miller Units)and the growth curve values are the average of two independant clones with their standard deviation that were measured during the same experiment. Experiment shows representative data which was obtained in the same way from three independent experiments.
The inducible promoter PliaI was also evaluated with the reporter vector pSBBs1C-lacZ which contains the lacZ reporter gene 
(Fig.2). Promoter activity leads to the expression of the β-galactosidase which directly correlates to the promoter activity. The β-galactosidase assay of the inducible Bacillus promoter PliaI was repeated three times. Data show one representative result.
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. </p>
