What is the Difference Between GFP and EGFP?
🆚 Go to Comparative Table 🆚The main differences between Green Fluorescent Protein (GFP) and Enhanced Green Fluorescent Protein (EGFP) are their sensitivity, brightness, and usage in molecular cloning:
- Definition: GFP is a wild-type green fluorescent protein used in molecular cloning of non-mammalian cells, while EGFP is an enhanced form of GFP that can be used on mammalian cells during molecular cloning.
- Origin: GFP is isolated from jellyfish (Aequorea victoria), whereas EGFP is synthesized in the laboratory by creating enhanced mutants.
- Sensitivity: GFP has low sensitivity, while EGFP has high sensitivity.
- Emission of fluorescence signals: GFP emits low fluorescence signals, while EGFP emits high fluorescence signals.
- Excitation peak: GFP has a major peak at 395 nm and a minor peak at 475 nm, while EGFP has a single excitation peak at 490 nm.
- Fluorescence at 37°C: GFP fluoresces less intensely than EGFP at 37°C.
- Folding efficiency at 37°C: GFP has low folding efficiency at 37°C, while EGFP has high folding efficiency at 37°C.
Both GFP and EGFP are used as reporter proteins in gene expression studies and can be synthesized using recombinant DNA technology. EGFP is a widely used variant of GFP, with mutations at two positions (F64L and S65T), making it brighter and able to mature rapidly at 37°C compared to wild-type GFP.
Comparative Table: GFP vs EGFP
Here is a table summarizing the differences between GFP and EGFP:
| Feature | GFP (Green Fluorescent Protein) | EGFP (Enhanced Green Fluorescent Protein) |
|---|---|---|
| Definition | Wild-type green fluorescent protein | Enhanced form of GFP |
| Origin | Isolated from jellyfish (Aequorea victoria) | Synthesized in the laboratory as an improved mutant |
| Sensitivity | Low | High |
| Emission | Major peak at 395 nm, minor peak at 475 nm (excitation) | Single excitation peak at 490 nm (excitation) |
| Uses | Molecular cloning of non-mammalian cells | Can be used on mammalian cells during molecular cloning |
| Fluorescence at 37°C | Less intense than EGFP at 37°C | More intense and quicker maturation at 37°C compared to GFP |
| Folding efficiency at 37°C | Low at 37°C | High at 37°C |
Both GFP and EGFP are used in molecular cloning and gene expression studies, where they function as reporter proteins. They are synthesized using recombinant DNA technology and can be further mutated to create improved forms.
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