A new bright green-emitting fluorescent protein--engineered monomeric and dimeric forms
Robielyn P Ilagan1, Elizabeth Rhoades, David F Gruber
1Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520, USA.
The FEBS Journal
|March 30, 2010
Summary
A new bright green fluorescent protein (VFP) from coral matures easily and is twice as bright as EGFP. Researchers engineered monomeric and dimeric VFP variants for diverse biological detection applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Fluorescent proteins are indispensable tools in biological research.
- Existing fluorescent proteins have limitations in brightness or stability.
Purpose of the Study:
- To discover and characterize a novel fluorescent protein from Cyphastrea microphthalma.
- To engineer variants with altered oligomerization states.
- To evaluate the performance of these variants compared to EGFP.
Main Methods:
- Isolation and characterization of vivid Verde fluorescent protein (VFP).
- Homology modeling and site-directed mutagenesis to alter oligomerization.
- Spectroscopic analysis and brightness comparison with EGFP.
- In vitro and in vivo application testing.
Main Results:
- VFP matures readily at 37°C and emits bright green fluorescence.
- Engineered monomeric (mVFP, mVFP1) and dimeric (dVFP) variants were created.
- VFP variants exhibit at least twice the brightness of EGFP.
- Demonstrated effectiveness in both in vitro and in vivo detection.
Conclusions:
- Vivid Verde fluorescent protein (VFP) is a bright, stable fluorescent protein with potential for various applications.
- Engineered VFP variants offer tunable oligomerization states for specific experimental needs.
- VFP represents a valuable addition to the toolkit of fluorescent proteins for biological imaging and detection.
