Team:Uppsala University/Promoters

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OBS! PRELIMINÄRT
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The promoter test was conducted by measuring florescence expressed by each individual promoter assembled with B0032-SYFP2 (Bba_**) though a FACS (florescence activated cell sorter) in lab strains MG1655 and D5Alpha.
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The test was conducted on triplicated cultures of each promoter construct, which were measured in the FACS. For each culture 105 cells fluorescence were measured individually, dead cells and non fluorescent cells were discarded, and a average where calculated. All promoter constructs of each strain were grown under same conditions.
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The variance in expression between MG1655 and D5Alpha may depend on the J23101-B0032-SYFP2, which is the promoter the other promoters strength is calculated from, construct in D5Alpha might been a weaker phenotype resulting in higher RPU values to other D5Alpha
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Alternatively the maximum protein expression is lower in D5Alpha due to its lower fitness also resulting in lower expression of SYFP2 in the J23101 construct.
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Revision as of 23:52, 25 September 2012

Team Uppsala University – iGEM 2012


OBS! PRELIMINÄRT The promoter test was conducted by measuring florescence expressed by each individual promoter assembled with B0032-SYFP2 (Bba_**) though a FACS (florescence activated cell sorter) in lab strains MG1655 and D5Alpha. The test was conducted on triplicated cultures of each promoter construct, which were measured in the FACS. For each culture 105 cells fluorescence were measured individually, dead cells and non fluorescent cells were discarded, and a average where calculated. All promoter constructs of each strain were grown under same conditions. The variance in expression between MG1655 and D5Alpha may depend on the J23101-B0032-SYFP2, which is the promoter the other promoters strength is calculated from, construct in D5Alpha might been a weaker phenotype resulting in higher RPU values to other D5Alpha Alternatively the maximum protein expression is lower in D5Alpha due to its lower fitness also resulting in lower expression of SYFP2 in the J23101 construct.



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