Difference between revisions of "Part:BBa K2926056"
 
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The extracellular domain of <i>S. cerevisiaes</i> membrane protein Opy2 was N-terminally fused to mCherry to enable protein visualization. Additionally, the major coat protein pVIII of the bacteriophage M13 was fused to the C-terminus of mCherry.
 
The extracellular domain of <i>S. cerevisiaes</i> membrane protein Opy2 was N-terminally fused to mCherry to enable protein visualization. Additionally, the major coat protein pVIII of the bacteriophage M13 was fused to the C-terminus of mCherry.
  
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===Usage and Biology===
 
===Usage and Biology===
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<div>This part was used during Troygenics Assembly to visualize the produced M13 bacteriophage-derived Troygenics via fluorescence measurement (Fig. 1).<br></div>
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<img class="figure image" style="width:800px" src="https://2019.igem.org/wiki/images/0/0b/T--Bielefeld-CeBiTec--Results_Troygenic_Assembly_final_fluorescence_spectrum.png">
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<figcaption> <b>Fig. 1: Emission spectrum of our mCherry-marked Toygenics with mCherry as a comparison measured with TECAN infinite M200 plate reader(λ[Ex]=570&nbsp;nm, λ[Em]=600&nbsp;nm to 850&nbsp;nm).</b></figcaption>
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We compared the emission spectrum of our Troygenics with the emission spectrum of mCherry, because the fluorescence marker cloned on the Troygenics is mCherry, too. We have seen a difference in the emission peak of about 20 nm. This is probably because the mCherry on our Troygenics is fused to the whole Troygenic. Because of this, there might be a different folding resulting in a shift of the emission spectrum.
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Latest revision as of 12:40, 21 October 2019


Fusion protein of Opy2 from yeast, mCherry and pVIII from the bacteriophage M13

The extracellular domain of S. cerevisiaes membrane protein Opy2 was N-terminally fused to mCherry to enable protein visualization. Additionally, the major coat protein pVIII of the bacteriophage M13 was fused to the C-terminus of mCherry.


Usage and Biology

This part was used during Troygenics Assembly to visualize the produced M13 bacteriophage-derived Troygenics via fluorescence measurement (Fig. 1).
Fig. 1: Emission spectrum of our mCherry-marked Toygenics with mCherry as a comparison measured with TECAN infinite M200 plate reader(λ[Ex]=570 nm, λ[Em]=600 nm to 850 nm).
We compared the emission spectrum of our Troygenics with the emission spectrum of mCherry, because the fluorescence marker cloned on the Troygenics is mCherry, too. We have seen a difference in the emission peak of about 20 nm. This is probably because the mCherry on our Troygenics is fused to the whole Troygenic. Because of this, there might be a different folding resulting in a shift of the emission spectrum.


Sequence and Features

Sequence was validated by Sanger sequencing.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]