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

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The strong constitutive promoter-EYFP generator (<partinfo>BBa_K804001</partinfo>) includes an enhanced yellow fluorescence protein  (<partinfo>BBa_E0030</partinfo>) under the control of a constitutive pTet promoter (<partinfo>BBa_J23118</partinfo>). This composite part's purpose is to constitutively express the YFP(<partinfo>BBa_E0030</partinfo>), which is also available with a strong ribosome binding site (<partinfo>BBa_B0034</partinfo>).
 
The strong constitutive promoter-EYFP generator (<partinfo>BBa_K804001</partinfo>) includes an enhanced yellow fluorescence protein  (<partinfo>BBa_E0030</partinfo>) under the control of a constitutive pTet promoter (<partinfo>BBa_J23118</partinfo>). This composite part's purpose is to constitutively express the YFP(<partinfo>BBa_E0030</partinfo>), which is also available with a strong ribosome binding site (<partinfo>BBa_B0034</partinfo>).
  
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<p align=center>https://static.igem.org/mediawiki/2012/2/29/UBCYFPCultures.png</p> '''Fluorescence Output of EYFP under a pTet constituitive promoter in co-culture and mono-culture :''' MetA- auxotroph is transformed with the EYFP construct (<partinfo>BBa_K804001</partinfo>) under the constitutive pTet promoter (<partinfo>BBa_J23118</partinfo>). We co-cultured this with TyrA- auxotrophs containing a ECFP construct (<partinfo>BBa_K804000</partinfo>)under the same constitutive pTet promoter; as well as with TrpA- auxotrophs containing a RFP construct (<partinfo>BBa_K081012</partinfo>) under the same constitutive pTet promoter. 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.
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<p align=center>https://static.igem.org/mediawiki/2012/2/29/UBCYFPCultures.png</p> '''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 (<partinfo>BBa_K804001</partinfo>) under the constitutive pTet promoter (<partinfo>BBa_J23118</partinfo>). We co-cultured this with TyrA- auxotrophs containing a ECFP construct (<partinfo>BBa_K804000</partinfo>)under the same constitutive pTet promoter; as well as with TrpA- auxotrophs containing a RFP construct (<partinfo>BBa_K081012</partinfo>) 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.
  
 
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Revision as of 01:50, 4 October 2012

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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.


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