Part:BBa_K118011:Experience
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Applications of BBa_K118011
Preliminary tests of this part were conducted using the reporter gene xylE (BBa_K118021). Strong repression occurred in the presence of glucose, and partial repression in the presence of high concentrations of higher sugars. Results can be found [http://2008.igem.org/Team:Edinbrugh/Results/PcstA-xylE here].
Characterization of PcstA with RFP Generator Part:BBa K081014
User Reviews
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BBa_K118011 1 Not understood Kun |
Cells with BBa_K200018 were grown overnight in various different media, and the GFP fluorescence was measured. |
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BBa_K118011 2 Not understood WHU-China 2012 |
Firstly,by embedding the gene of RFP downstream the promoter PcstA,we have successfully certified that the promoter PcstA can be activated by CRP.In other words, it can be repressed by high concentration of glucose. What's more, we have constructed a indirect pathway by using an intermediate to change the function of the promoter. In the indirect pathway,PcstA is activated by high concentration of glucose. After construction,the correct clones were cultured in 96-well plate at 37℃ for 24 hours,then the fluorescence and absorbance at 600 nm were recorded on a SpectraMax M2 plate reader.All fluorescence was normalized with absorbance at 600 nm.The cell density was characterized by the absorbance at 600nm,which increased with glucose concentration.The result is showed in the figure bellow.
In this figure, the fluorescence of RFP generator promoted by PcstA decreased when concentration of glucose rose, meanwhile, the fluorescence of the indirect regulatory device increased with the glucose concentration. In another word, the indirect pathway can be activated by glucose For more information, please go to wiki of WHU-China 2012 on [http://2012.igem.org/Team:WHU-China/Project?catalog=2#Indirect_Pathway_Design Indirect pathway] |
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