Difference between revisions of "Part:BBa K2926054"
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Mating factor alpha was N-terminally fused to the red fluorescent protein mCherry. Additionally the major coat protein pVIII of the bacteriophage M13 was fused to the N-terminus of mCherry. | Mating factor alpha was N-terminally fused to the red fluorescent protein mCherry. Additionally the major coat protein pVIII of the bacteriophage M13 was fused to the N-terminus of mCherry. | ||
+ | ===Usage and Biology=== | ||
+ | <html> | ||
+ | This part was used during Troygenics Assembly to visualize the produced M13 bacteriophage-derived Troygenics via fluorescence measurement (Fig. 1).<br> | ||
+ | <div class="middle"> | ||
+ | <figure class="figure large"> | ||
+ | <img class="figure image" src="https://2019.igem.org/wiki/images/0/0b/T--Bielefeld-CeBiTec--Results_Troygenic_Assembly_final_fluorescence_spectrum.png"> | ||
+ | <figcaption>Fluorescence of a cultures of <i>E. coli</i> Fig. 1: Emission spectrum of our mCherry-marked Toygenics with mCherry as a comparison measured with TECAN infinite M200 plate reader(λ<sub>Ex</sub>=570 nm, &lambda<sub>Em</sub>=600 nm to 850 nm).</figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | <div> | ||
+ | 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. | ||
+ | </div> | ||
+ | |||
+ | </html> | ||
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Revision as of 12:05, 21 October 2019
Fusion protein of mating factor alpha from yeast, mCherry and pVIII from the M13 bacteriophage
Mating factor alpha was N-terminally fused to the red fluorescent protein mCherry. Additionally the major coat protein pVIII of the bacteriophage M13 was fused to the N-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).
![](https://2019.igem.org/wiki/images/0/0b/T--Bielefeld-CeBiTec--Results_Troygenic_Assembly_final_fluorescence_spectrum.png)
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 via Sanger sequencing.
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]