Difference between revisions of "Part:BBa K4335002"
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</figcaption> | </figcaption> | ||
</figure> | </figure> | ||
+ | <figure> | ||
+ | <img src="https://static.igem.wiki/teams/4335/wiki/staygoldfzx1.png" width="75%" style="float:center"> | ||
+ | <figcaption> | ||
+ | <p style="font-size:1rem">StayGold and other 15 kinds of fluorescent protein fluorescence intensity changes with time | ||
+ | </p> | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | |||
+ | StayGold has an order of magnitude more light stability than any currently available fluorescent protein and has a cell brightness similar to that of mNeonGreen. Compared with the 15 fluorescent proteins, StayGold showed good photostability. | ||
+ | |||
+ | <h2>Usage</h2> | ||
+ | We introduced the StayGold gene into the <a href="https://parts.igem.org/Part:BBa_K4335040">[pTX2040]</a> plasmid. | ||
+ | PCrRBCS2 is used as the promoter and TCrRBCS2 is used as the terminator to achieve the expression of the StayGold gene. | ||
+ | <h2>Result</h2> | ||
+ | <h3>Plasmid construction</h3> | ||
+ | We designed two primers: Staygold-F and Staygold-R, and confirmed the successful assembly by PCR amplification. | ||
+ | <figure> | ||
+ | <img src="https://static.igem.wiki/teams/4335/wiki/staygoldfzx4.png" width="100%" style="float:center"> | ||
+ | <figcaption> | ||
+ | <p style="font-size:1rem"> | ||
+ | </p>The primers | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | |||
+ | <figure> | ||
+ | <img src="https://static.igem.wiki/teams/4335/wiki/staygoldfzx2.png" width="100%" style="float:center"> | ||
+ | <figcaption> | ||
+ | <p style="font-size:1rem">Primers targeted location | ||
+ | </p> | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | |||
+ | <figure> | ||
+ | <img src="https://static.igem.wiki/teams/4335/wiki/staygoldfzx3.png" width="20%" style="float:center"> | ||
+ | <figcaption> | ||
+ | <p style="font-size:1rem">The electrophoretogram of StayGold after amplification | ||
+ | </p> | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | |||
+ | <h3>Functional Identification</h3> | ||
+ | |||
+ | <figure> | ||
+ | <img src="https://static.igem.wiki/teams/4335/wiki/staygoldfzx5.jpg" width="100%" style="float:center"> | ||
+ | <figcaption> | ||
+ | <p style="font-size:1rem">Fluorescence excitation of positive clones transferred into pTX2040 vector compared with wild-type mCherry, together with DIC field and chlorophyll excitation as fluorescence controls. | ||
+ | </p> | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | <h2>Reference</h2> | ||
+ | [1] Hirano, M. et al. A highly photostable and bright green fluorescent protein. Nat. Biotechnol. 40, 1132–1142 (2022). | ||
</html> | </html> | ||
Revision as of 08:22, 11 October 2022
StayGold, a newly discoverd green fluorescent protein
StayGold, a green fluorescent protein (GFP) , fluoresces in expression. StayGold has an order of magnitude more photostability than any currently available fluorescent protein. Commonly used to label and trace cellular components. BBa_K4335002 is our StayGold gene optimized for the codon of Chlamydomonas reinhardtii.
Additional Materials
StayGold was extracted from Cytaeis uchidae by Atsushi Miyawaki et al. by the RIKEN Center for Brain Science Research on April 25, 2022. StayGold has an order of magnitude more light stability than any currently available fluorescent protein and has a cell brightness similar to that of mNeonGreen. Compared with the 15 fluorescent proteins, StayGold showed good photostability.Usage
We introduced the StayGold gene into the [pTX2040] plasmid. PCrRBCS2 is used as the promoter and TCrRBCS2 is used as the terminator to achieve the expression of the StayGold gene.Result
Plasmid construction
We designed two primers: Staygold-F and Staygold-R, and confirmed the successful assembly by PCR amplification.Functional Identification
Reference
[1] Hirano, M. et al. A highly photostable and bright green fluorescent protein. Nat. Biotechnol. 40, 1132–1142 (2022).Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 540
- 1000COMPATIBLE WITH RFC[1000]