Green Fluorescent Protein

In the last few years, green fluorescent protein (GFP) has become one of the most widely used tools in molecular genetics. The fact that GFP can generate an internal highly visible fluorophore has made it a tremendously valuable marker or reporter gene which can be utilised in virtually every field of molecular cell genetics. One of the problems associated with this method is how to confirm that the gene of interest is being either expressed or activated in the target cells and to what extent this activation or expression occurs. The use of conventional reporter genes such as luciferase or lacZ require that the cells be processed and/or treated with substrates in order to monitor the reporter gene. The fact that GFP requires no additional substrates and can therefore be montiored non-invasively in living cells has been a revolutionary advance. In addition, a number of mutations in the wild type GFP protein, mean that it can also be used as a quantitative marker in living cells. GFP can also be used as a tag for studying localisation and movement of protein-GPF chimeras within cells in real time. There are also a growing number of GFP mutants with different spectral properties which allow, for example, simultaneous monitoring of proteins labelled with different GFPs or the use of fluorescent energy transfer techniques to measure protein-protein interations.