Final answer:
Chromatography, specifically affinity chromatography, is used to purify GFP from other bacterial proteins by exploiting GFP's unique affinity to a specific ligand attached to a chromatography column. Other proteins are washed away, and GFP is then eluted, identified by its green fluorescence under UV light.
Step-by-step explanation:
Purification of Green Fluorescent Protein (GFP)
Chromatography is a method used to purify proteins such as GFP from a mixture of other bacterial proteins. One common type of chromatography used is affinity chromatography, which exploits the unique properties of GFP. In this process, a column with a resin is used that has a ligand specific to GFP. When the bacterial lysate is passed through the column, GFP binds to the ligand due to its affinity, while other proteins do not.
Once bound, the column can be washed to remove the unwanted proteins. After washing, GFP is eluted by adding a solution that competes with GFP for binding to the ligand or that changes the conditions so that GFP no longer binds to the column. Thus, GFP is separated from the rest of the bacterial proteins. The fluorescence property of GFP can also facilitate its detection and quantification post-purification, often by using UV light to observe its characteristic green fluorescence.
Different chromatography techniques can be tailored according to the properties of the target protein and contaminants for efficient separation, such as size, charge, hydrophobicity, or specific binding affinity.