Difference between revisions of "Part:BBa K143012:Experience"

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We wanted to work with this promoter and evaluate it as part of our Bacillus BioBrickBox, but the DNA was not available. So we created a new BioBrick and characterized it with the reporter ''lux'' operon.
 
We wanted to work with this promoter and evaluate it as part of our Bacillus BioBrickBox, but the DNA was not available. So we created a new BioBrick and characterized it with the reporter ''lux'' operon.
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===Evaluation===
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[[Image:Englisch Auswertung PliaG Pveg.png|thumb|right|400px| <p align="justify"> '''β-galactosidase assay and growth curve of strains carrying the promoters P<sub>''liaG''</sub> (black) and P<sub>''veg''</sub> (grey) 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"> The two constitutive promoters P<sub>''liaG''</sub> and P<sub>''veg''</sub> were evaluated with the reporter ''lacZ''. 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 P<sub>''veg''</sub> and P<sub>''liaG''</sub> 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 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 12:03, 25 September 2012

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K143012

User Reviews

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Korinna

We wanted to work with this promoter and evaluate it as part of our Bacillus BioBrickBox, but the DNA was not available. So we created a new BioBrick and characterized it with the reporter lux operon.

Evaluation

β-galactosidase assay and growth curve of strains carrying the promoters PliaG (black) and Pveg (grey) 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 two constitutive promoters PliaG and Pveg were evaluated with the reporter lacZ. 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. 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.

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Chris

This part was used as part of a promoter and RBS pair (BBa_K143053) as part of the Biofabricator subtilis Imperial iGEM 2008 project. When incorporated into GFP and RFP production constructs(BBa_K143079 and BBa_K143082), the promoter and RBS pair successfully produced a GFP or RFP output.

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