Difference between revisions of "Part:BBa K5317002"
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | The monomeric near-infrared (NIR) fluorescent protein miRFP670 was developed | + | The monomeric near-infrared (NIR) fluorescent protein miRFP670 was developed by Shcherbakova and colleagues in 2016. |
− | The | + | The excitation maxima of miRFP670 is at 642 nm and the emission maxima at 670 nm. miRFP670 was genetically engineered from bacterial phytochrome photoreceptors (BphP) to generate reporters using the optical window with low autofluorescence, decreased light scattering, and minimal absorption of biological substances such as hemoglobin, water, and melanin. Of the BphP-based fluorophores, miRFP is particularly suitable for investigations in mammalian cell lines due to a fluorescence quantum yield of 14 % and maintaining brightness in the physiological pH range from 5 to 8 (Shcherbakova ''et al.'', 2016). |
− | miRFP670 was employed in our mammalian cell-based antibiotic detection sensor downstream of the ATF2- | + | miRFP670 was employed in our mammalian cell-based antibiotic detection sensor downstream of the ATF2-3xCre3xAP1 promoter (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K5317017 K5317017]</span>) visualizing the antibiotic presence-dependent phosphorylation of ATF2 (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K5317016 K5317016]</span>). |
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Latest revision as of 21:22, 1 October 2024
miRFP670
Usage and Biology
The monomeric near-infrared (NIR) fluorescent protein miRFP670 was developed by Shcherbakova and colleagues in 2016. The excitation maxima of miRFP670 is at 642 nm and the emission maxima at 670 nm. miRFP670 was genetically engineered from bacterial phytochrome photoreceptors (BphP) to generate reporters using the optical window with low autofluorescence, decreased light scattering, and minimal absorption of biological substances such as hemoglobin, water, and melanin. Of the BphP-based fluorophores, miRFP is particularly suitable for investigations in mammalian cell lines due to a fluorescence quantum yield of 14 % and maintaining brightness in the physiological pH range from 5 to 8 (Shcherbakova et al., 2016).
miRFP670 was employed in our mammalian cell-based antibiotic detection sensor downstream of the ATF2-3xCre3xAP1 promoter (K5317017) visualizing the antibiotic presence-dependent phosphorylation of ATF2 (K5317016).
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 763
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 763
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 419
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 763
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 763
Illegal NgoMIV site found at 82
Illegal NgoMIV site found at 370
Illegal NgoMIV site found at 405 - 1000COMPATIBLE WITH RFC[1000]
Reference
Shcherbakova, D. M., Baloban, M., Emelyanov, A. V., Brenowitz, M., Guo, P., & Verkhusha, V. V. (2016). Bright monomeric near-infrared fluorescent proteins as tags and biosensors for multiscale imaging. Nature communications, 7, 12405. https://doi.org/10.1038/ncomms12405