The green fluorescent protein has gained significant attention in biology, medicine and research and has been described as the microscope of the twenty first century for a very good reason. Through this protein, it has become easy to not only observe proteins as they are being made, but also observe any movements. By attaching the gene of this protein to the gene of a given protein or an organism, scientists and researchers can easily observe any protein of interest given that GFP fluoresces.
Because it can be attached to other proteins and organisms, GFP has become one of the most popular imaging tools. Proteins in particular are very small and can prove very difficult to observe. However, by attaching GFP to the protein (as a tag) the green fluorescence of the protein enables the protein of interest to be viewed. It is for this reason that GFP is referred to as the modern microscope.
For a long time, neuroscientists were unable to activate/stimulate single neurons given that they could only stimulate the brain cells with electrodes. However, through optogenetics, it is now possible to stimulate individual neurons instantly. This is achieved by using an algae protein (attached to the neuron of interest) as well as light.
Here, the fluorescent protein is used to indicate which of the neurons has been manipulated to become the on and off switch.
Today, a good number of studies in this field have been directed towards understanding the photochemistry of the protein (GFP) and using its model for the purposes of mimicking its chromophore. This has helped in the development of DMFBI-RNA complex referred to as Spinach.
is a fluorescent RNA tag that is selective and non-toxic. The compound only
fluoresces when it is attached to an RNA, which means that it can help follow the
molecules of interest as they move through cells. Apart from being non-toxic,
this RNA tag is also resistant to photo bleaching, which means that it provides a great service.
By using a Brainbow of colors, it has become possible for researchers to map neural circuits of the brain. For instance, researchers used this method to introduce genetic machinery, which randomly mixes green, cyan and yellow fluorescent proteins in various individual neurons to create a palette of 90 distinctive hues of colors.
Through this technique, it is now also possible to differentiate between given neurons and learn more about them. For instance, by labeling say 100 neurons in a single mouse, this becomes more convenient and efficient than labeling a single neuron in 100 mice for studies.
Looking for fluorescence through microscopy is one of the ways through which GFP fusion expression can be assessed. This approach has been successfully used to show that a GFP moiety has been accurately expressed making a straightforward method of assessing fusion expression.
It may prove difficult to prove that an entire fusion
protein is being made through microscopy. For this reason, the immunoblot method is
largely preferred given that it allows for the detection of both immature and
The discovery and understanding of GFP has also made it possible to make good observations of such organisms as yeast. Given a good number of yeast media yellow auto fluorescence once they are excited by a specific light (UV or blue light) fluorescence filters that maximize the detection of GFP (while at the same time minimizing auto fluorescence) have to be used.
Today, GFP is being extensively used in many experiments making it a very important scientific tool. Because of its strengths, it has proved to be very important for studying the dynamics of various proteins, nucleic acids as well as lipid localization in yeast.
In addition, it is a beneficial tool for studying the movement and functions of various cell organelles, interaction of proteins, gene expression as well as studying the structure of proteins among many other uses. Therefore, it can be argued that GFP has become an important tool in biology, medicine and research, making significant contributions to microscopy.
Marcy Patrick (2014) Plasmids 101: Green Fluorescent Protein (GFP)
Marc Zimmer (2015) Green Fluorescent Protein: A Molecular Microscope.
Martin Chalfie and Steven R. Kain (2005) Green Fluorescent Protein: Properties, Applications and Protocols.
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