Fluorescence is the process by which the fluorophore absorbs a stimulus like light on interaction. This causes a conformational change in the fluorophore where a longer wavelength is created via an energy transfer process. The lower emission of photons through the change can be detected as an electrical signal. This phenomena can be seen in aromatic biological proteins like tryptophan and tyrosine through the imidazole ring, allowing them to be synthesized in an array of environments Williams, Slatko and McCarrey, 2007). Fluorescence was first discovered in Aequorea victoria, which synthesizes aquorin, a chemiluminescent protein. It is a luciferase that catalyses coelenterazine oxidation through a calcium dependent reaction to emit blue light and green fluorescence under UV light. Soon after, it was cloned and GFP was expressed in bacteria (Williams, Slatko and McCarrey, 2007). Luciferase is a bacterial reporter found in some species like the Photobacterium. The bioluminescence is produced from the dehydration of hydroflavin intermediate, producing blue/green light. Mutations were discovered that enhanced the efficiency of GFP -giving rise to enhanced fluorescent proteins(EGFP). The point mutation could then be used in mammalian cells due to increased stability. Likewise other mutant colour variants were created from homologues of GFP like the marine corals, which revolutionized the understanding of disease and many biological processes. The variances allows the use of colour due to the unique β barrel formation of FP. However, this can also adversely affect the brightness and the sensitivity too (Kremers et al., 2011). One of the first uses of FP was in protein labeling. This allowed the expression and localisation of prot... ... middle of paper ... ...erm. It also causes hypersensitisation and a relocalisation of fluorescence within the tissue. It is thought that this relocalisation simply due to a change in affinity for the living and dead cells. With prolonged use of photosensitization, the morphology of the cells can change (Serebrovskaya et al., 2009). The practical implication of the green fluorescent proteins is extensive, affecting the study at a genomic level all the way to an organismal level. It has greatly impacted on all these areas of research with a vast array of techniques now available all based on this concept. The potential of FPs was quickly realized and many of the limiting features have been used at an advantage as seen with FRAP. Whilst many of the conventional uses were to allow visualization of molecules/subcellular compartments, FP can be used in drug testing and as medication itself.
Enhanced green fluorescent protein (EGFP) was originally isolated from a bioluminescent jellyfish called Aequorea victoria. As suggested by the name, this protein fluoresces green when exposed to light in the ultraviolet range. The ultimate goal of the following experiment was to successfully create a pET41a(+)/EGFP recombinant plasmid that was transformed into live E. coli cells. The success of this transformation could be evaluated based on whether EGFP’s fluorescence properties were displayed by the colony in question. The protein’s fluorescence properties “triggered the widespread and growing use of GFP as a reporter for gene expression and protein localization in a broad variety of organisms” (Ormo, et. al., 1996). Although EGFP and GFP differ for a few amino acids that make EGFP’s fluorescence mildly stronger, the basic principle that such a protein allows for the evaluation of transformation success remains intact.
Serratia marcescens, a Gram-negative bacillus, was originally and solely considered a biological marker in the medicinal industry, due to its highly natural red pigment: Prodigiosin (Hejazi and Falkiner, 1997). The pigment has numerous roles within bacteria, which can be further translated into the pharmaceutical and medical domain. This bacterium naturally occurs in water, soil, on plants as well as in humans and animals (Khanafari et al, 2006), where it is deemed an opportunistic pathogen.
al. (1994) explain that a complementary DNA for GFP produces a fluorescent product when expressed in E. coli cells as the expression of GFP can be used to monitor gene expression and protein localization in living things. In this experiment, the heat shock method will be used to deliver a vector (plasmid) of GFP to transform and grow E. coli bacteria. Four plates containing Luria Bertani (LB) broth and either –pGLO or +pGLO will have E. coli bacteria added to it. The plate containing –pGLO (no pGLO) and LB will show growth as ampicillin will be present killing bacteria but no glowing because no arabinose will be present for glowing to be activated, the same result will be seen in the plate containing +pGLO, LB and ampicillin.
The plasmid pGLO contains the GFP gene which can only be activated while in the presence of arabinose. The plasmid also contains an ampicillin resistance gene, which explains why bacterial cells transformed with pGLO can survive when ampicillin is present. GFP is a protein that is found in jellyfish, and is known for emitting a green fluorescent light. In vitro, it is able to fluoresce this green color by releasing photons from being in an excited energy state. The energy was provided by a UV light at the end of this experiment. Jellyfish can fluoresce this green on their own because in vivo, GFP works with another protein that allows the GFP to fluoresce without being in an excited energy
1. In response to light, phytochrome undergoes a change in shape that leads to the activation of
Sherin, Jonathan E., and Charles B. Nemeroff. "Abstract." National Center for Biotechnology Information. U.S. National Library of Medicine, 02 Mar. 0006. Web. 11 Dec. 2013.
Albinism is a genetic condition present at birth, characterized by a small amount of melanin pigment in the skin, hair and eye. Albinism is an occasional inborn sickness related with vision difficult, which affect one in seventeen thousand persons. It is not a contagious disease and cannot be spread over contact. Albinism affects individuals from all races. Most folks with albinism have parents with a normal color of skin. Some may not even recognize that they are Albino until later on in their life. This paper will be based on the study of albinism, causes, types, the genetic transmission and some possible medical problem.
...he red and far-red light through photo-interconversion between the two stable conformations. Chromophores are responsible in mediated this distinct feature (Blumenstein et al., 2005). A. niger also produce fumonisins, one of the well-known industrial workhorses (Dufossé, et al., 2014).
The absorption of light in the form of photons through the thylakoid membrane into the lumen is the first step of photosynthesis. This photons absorbed through the lumen go through photochemical reduction in which they are absorbed into pigments such as chlor...
This experiment synthesized luminol (5-Amino-2,3-dihydro-1,4-phthalazinedione) and used the product to observe how chemiluminescence would work. The starting material was 5-nitro-2,3-dihydrophthalazine-1,4-dione, which was, after addition of reaction agents, refluxed and vacuum filtered to retrieve luminol. Using two stock solutions, we missed our precipitated luminol with sodium hydroxide, potassium ferricyanide, and hydrogen peroxide, in their respective solutions, in a dark room, to observe the blue light
Fulda, K. G., and K. Lykens. "Abstract." National Center for Biotechnology Information. U.S. National Library of Medicine, 25 Aug. 0005. Web. 18 Mar. 2014. .
Devlin, Robert M. and Allen V. Barker. Photosynthesis. New York, Van Nostrand Reinhold Company, 1971.
Photosynthetic pigments are essential for life because they allow photosynthesis to occur by capturing sunlight which is then used alongside carbon dioxide and water to form organic compounds such as glucose and oxygen. The pigments allow the conversion of light energy to chemical energy which other organisms can benefit from. Oxygen is utilised by other organisms in aerobic respiration. The different pigments present in the chloroplasts allow a wide variety of wavelengths of light to be absorbed for efficient photosynthesis and provide colours to the plant to attract pollinators.
The initial motivation for scientists to investigate more about dye ligand affinity chromatography was given after the interactions that took place between Blue Dextran, a Cibaron Blue and dextran conjugate, which is used as a void marker in size-exclusion chromatography, and particular kinases. Until then, only purification of various proteins by size-exclusion chromatography with Blue Dextran, like for example, erythrocyte pyruvate kinase, phosphofructokinase, glutathione reductase, and several coagulation factors had been initiated. The final conclusion of these studies was that the main reason for protein binding was Cibacron Blue F3G-A, a major reactive dye (Denizli and Pişkin, 2001).
In plants, proteins called photosynthetic reaction centers contain green chlorophyll that absorbs light energy. These proteins are held inside organelles called chloroplasts, which is abundant in leaf cells. In contrast, bacteria house the proteins in the plasma membrane. Chloroplasts are found in the cells of green plants and photosynthetic algae where photosynthesis takes place. Inside the chloroplast are folded structures in disk-shaped arrangement called thylakoids, which enclose chlorophyll in their membrane. Only certain portions of the light spectrum can be absorbed and the photosynthetic action spectrum is dependent on the type of accessory pigment present. Green plants mostly absorb red and blue wavelengths because the action spectrum corresponds to absorption spectrum for chlorophylls and carotenoids. The color of the pigment comes from the wavelengths of light reflected. Plants appear green because they reflect yellow and green wavelengths of light. Photosynthesis involves two series of chemical events, called the light independent that occurs in the stroma and light dependent reactions that occurs in the lumen. They are also known as light and dark reactions this terminology is somewhat ambiguous, because the entire process of photosynthesis is regulated to take place when an organism absorbs visible light. Organized clusters of chlorophyll and beta-carotene in the thylakoid membrane are present to