Part:BBa_E0030:Experience

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Characterization by British Columbia iGEM 2012

YFP fluorescence output by British Columbia iGEM 2012

The strong constitutive promoter-EYFP generator (BBa_K804001) includes an enhanced yellow fluorescence protein (BBa_E0030) under the control of a constitutive pTet promoter (BBa_J23118). This composite part's purpose is to constitutively express the YFP(BBa_E0030), which is also available with a strong ribosome binding site (BBa_B0034).

Fluorescence Output of EYFP under a pTet constituitive promoter in co-culture and mono-culture with MetA- auxotroph grown over time : MetA- auxotroph is transformed with the EYFP construct (BBa_K804001) under the constitutive pTet promoter (BBa_J23118). We co-cultured this with TyrA- auxotrophs containing a ECFP construct (BBa_K804000)under the same constitutive pTet promoter; as well as with TrpA- auxotrophs containing a RFP construct (BBa_K081012) under the same constitutive pTet promoter. These cells were grown in minimal media spiked with 10^-3M amino acids (methionine, tryptophan, and tyrosine) . Fluorescence output of EYFP for the three member co-culture, pairwise co-culture, and mono-culture with MetA- auxotrophs are analyzed using a plate reader which measures emission of YFP at a wavelength of 527nm when excited at 514nm.

User Reviews

UNIQf84b4476ea9a4ecf-partinfo-00000009-QINU UNIQf84b4476ea9a4ecf-partinfo-0000000A-QINU

••••• 2012 iGEM UIUC

The part worked as expected. The way we quantified it through a plate reader with emmission wavelengths. Excitation: 515nm Emission: 528nm We used this part for this project, data collected can be found http://2012.igem.org/Team:UIUC-Illinois/Results.

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