Difference between revisions of "Part:BBa K4335002"
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Revision as of 03:35, 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.
Absorption spectrum and emission spectrum of StayGold.[Image origin]
StayGold and other 15 kinds of fluorescent protein fluorescence intensity changes with time
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.
The primers
Primers targeted location
The electrophoretogram of StayGold after amplification
Functional Identification
Fluorescence excitation of positive clones transferred into pTX2040 vector compared with wild-type mCherry, together with DIC field and chlorophyll excitation as fluorescence controls.
Reference
[1] Hirano, M. et al. A highly photostable and bright green fluorescent protein. Nat. Biotechnol. 40, 1132–1142 (2022).Sequence and Features
- 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]