Exploring Circular Permutation of Red Fluorescent Proteins
Author Information
Author(s): Shui Bo Wang, Qi Lee, Frank Byrnes, Laura J. Chudakov, Sergey A. Lukyanov, Holger Sondermann, Michael I. Kotlikoff
Primary Institution: Cornell University
Hypothesis
Can circular permutation of mCherry and mKate produce bright and stable fluorescent proteins?
Conclusion
The study identified several highly fluorescent circularly permuted variants of mCherry and mKate, which can be used as building blocks for molecular sensors.
Supporting Evidence
- Four cp-mKates were identified with over 80% of native fluorescence.
- cp-mCherry194-193 retained 60.6% of the brightness of native mCherry.
- cp-mKate189-188 is the brightest circular permutation variant in E. coli.
- Structural analysis revealed that the circularly permutated proteins maintained similar structures to wild-type.
Takeaway
Scientists changed the order of parts of two glowing proteins to make new versions that still glow brightly, which can help in creating better sensors.
Methodology
The study involved creating tandem fusion templates of mCherry and mKate, screening for fluorescence in E. coli, and evaluating the brightness of various circular permutants.
Limitations
The fluorescence of some circularly permuted proteins was lower than expected, indicating incomplete folding.
Digital Object Identifier (DOI)
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