Research Project : Fluorescent Proteins And Characterisation Of The Citrine Gene And Yfp Produced

992 Words Mar 22nd, 2016 4 Pages
This research project aimed to utilise tools and techniques of molecular research to carry out cloning, expression and characterisation of the Citrine gene and YFP produced. Fluorescent proteins including GFP initially isolated from Aequorea victoria jellyfish have found many applications including a reporter for gene expression studies and a fluorescent probe for live cell imaging without potentially harmful exogenous substrates, especially to study the localization and dynamic behaviours of proteins (Shaner, Steinbach and Tsien, 2005). The early GFPs were quickly recognised to have far reaching potential despite these significant deficiencies (Heim, Cubitt and Tsien, 1995). The main disadvantages included pH sensitivity and dimerization, in dimerization, proteins agglutinate together affecting the characteristics of the fusion protein. Effort has been targeted in the improvement of fluorescent protein capabilities. Leading to the development of YFPs including Citrine and Venus, enabling fluorescent labellings not previously possible (Nagai et al., 2002). A wide range of fluorescent proteins are now available spanning across the visible spectrum, each with remarkably different properties and characteristics suitable for a variety of purposes (Figure 1). Relatively recent techniques through site-directed or random mutagenesis led to the regular development of new GFP variants with a range of excitation and emission wavelengths, increased brightness due to the quantum yield,…

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