GFP Recombinant Proteins

Green Fluorescent Protein (GFP) is a naturally occurring protein derived from the jellyfish Aequorea victoria. It exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. GFP recombinant proteins are widely used as molecular markers in biological research due to their ability to visualize and track cellular processes in real-time.

Production Process

• Gene Cloning: The gene encoding GFP is cloned into an expression vector tailored for the chosen host system, typically Escherichia coli or mammalian cells.
• Expression in Host Cells: The recombinant vector is introduced into host cells, which are cultured under optimal conditions to express the GFP protein.
• Protein Purification: After cell lysis, GFP proteins are purified using techniques such as affinity chromatography, ion-exchange chromatography, and size-exclusion chromatography to achieve high purity.
• Protein Refolding and Formulation: If necessary, proteins are refolded to attain their functional conformation and formulated with stabilizers for storage and application.

Applications

• Research:
  • Cellular Imaging: Used to visualize and track proteins, organelles, and other cellular structures in live cells.
  • Gene Expression Studies: Employed as a reporter gene to monitor gene expression and regulation.
  • Protein-Protein Interactions: Utilized in assays such as FRET (Förster Resonance Energy Transfer) to study interactions between proteins.
• Biotechnology:
  • Biosensors: Engineered GFP variants are used in biosensors to detect changes in cellular conditions, such as pH or ion concentrations.
  • Synthetic Biology: GFP is used to create visual outputs in synthetic biology circuits and systems.
• Diagnostics:
  • Marker for Transfection: GFP is used as a marker to verify successful transfection and expression in cells.
  • Immunoassays: Employed in immunoassays to detect the presence of specific antigens or antibodies.

Advantages

• Non-Invasive Visualization: GFP allows for real-time, non-invasive visualization of cellular processes without the need for additional substrates or cofactors.
• High Sensitivity: The bright fluorescence of GFP provides high sensitivity in detecting and quantifying biological molecules.
• Versatility: GFP can be fused to other proteins, enabling the study of their localization, expression, and interactions within cells.
• Customization: Variants of GFP with different fluorescence spectra (e.g., YFP, CFP) allow for multi-color imaging and simultaneous tracking of multiple targets.

Recombinant GFP proteins are indispensable tools in molecular and cellular biology, enabling detailed visualization and analysis of dynamic processes in living organisms. Their high sensitivity, versatility, and ability to be genetically encoded make them essential for a wide range of research and diagnostic applications.