General information on EGFPprotein
Enhanced green fluorescent protein or EGFP or simply GFP is a protein that exhibit bright green fluorescence when exposed to light in the blue to ultraviolet range. The major advantage of this protein is that it can form internal chromophore whithout requiring any accessory enzymes/substrates, cofactors, gene products other than molecular oxygen. This protein is often used as reporter gene. EGFP can be expressed throughout a given organism, in organs of interest or selected cells. GFP is frequently introduced through transgenic techniques . Regardless, EGFP protein doesn’t affect the genome of the species or that of the offspring. GFP protein has been expressed in a wide range of species including bacteria, fungi, yeast, mammals and even human cells.
The GFP protein is often used as a reporter gene for different essays regarding environmental toxicity levels. Indeed, EGFP protein has been used to demonstrate toxicity levels of different chemicals such as phenol, triclosan, parabens ethanol and p-formaldehyde. The fluroresence is a way of showing how different pollutants can affect the host cells, Another variable that can be easily measured using EGFP protein is the density of cells. A major advantage of EGFP protein is that, depending on how it was introduced in the host cells, in can be heritable. This allows continued study of cells and the tissues where the protein is expressed. Although EGFP protein itself doesn’t interfere with biological processes, it can interact when fused to proteins of interest. Hence, in order to maintain the protein of interest unmodified, careful design of linkers is needed.
In terms of structure, EGFP protein has a soda-shape like structure. It contains a beta barrel that consists of eleven β-strands and a, alpha helix that runs through the center. The beta barrel contains the chromophore which is located in the middle. The alpha helix is covalently bonded to chromophore 4-(p-hydroxybenzylidene)imidazolidin-5-one (HBI). Five shorter alpha helices are located on the ends of the structure.