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.
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