Protocol for the Transfection of GFP

The use of Green Fluorescent Protein revolutionized molecular and cellular biology; it allowed researchers to visualize cells and proteins in ways never before possible. Its impact on scientific research was so profound that in 2008 Osamu Shimomura won the Nobel Prize in Chemistry for his discovery and isolation of GFP from fluorescent jellyfish.

Definitions

• A gene is the sequence of deoxyribonucleic acid in cells that is responsible for the production of a specific protein. A plasmid is a separate piece of DNA that contains a gene of interest such as GFP. Transfection is the method for introducing a plasmid into cells.

Methods

• Transfection typically involves mixing a DNA plasmid containing a gene such as GFP with lipids or salts. Cells are exposed to this solution, which creates holes in the cell membrane, allowing the plasmid into the cell where the gene can be expressed as protein. Commercial transfection kits are available, or the reagents can be made in a laboratory. The choice of transfection method depends on the type of cells and plasmids being used. The GFP is visualized and monitored with equipment that detects fluorescence.

Uses

• GFP is usually attached to a protein of interest for transfection. The Nobel Committee singled out the tracking of "nerve cell damage during Alzheimer's disease" and "how insulin-producing beta cells are created in the pancreas of a growing embryo" as examples.