Difference between revisions of "Part:BBa E0030:Experience"

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<I>2012 iGEM UIUC</I>
 
<I>2012 iGEM UIUC</I>
 
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Revision as of 03:10, 4 October 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_E0030

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).

UBCYFPCultures.png

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

UNIQa882a842ac780da8-partinfo-00000009-QINU UNIQa882a842ac780da8-partinfo-0000000A-QINU

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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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