<div>Expression and purification of the Green Fluorescent Protein via
immobilized metal ion affinity.</div><div>Olga D. Sieluzycka</div><div>School of Biological and Chemical Sciences (Fogg Building)</div><div>Queen Mary University of London Mile End Road</div><div>London E14NS UK</div><div><b>ABSTRACT:</b> The aim of the experiment is to express and purify
recombinant His-tagged Green Fluorescent Protein (GFP) protein in<i>Escherichia coli</i> (E.coli) using metal affinity chromatography.
GFP is an excellent trafficking tool in modern biochemistry. The protein
was expressed in E.Coli and introduced to the required organism by a
plasmid transformation. Plasmid containing the GFP was controlled by the
T7 promoter had ampicillin resistance and had an N-terminal His6 tag
fusion to facilitate the process of purification in the later stages of
experiment. The expression of proteins was confirmed by the SDS gel by
comparing the bands widths. The goal of experiment was not fully met.</div><div><b>Introduction:</b> The main objective of the experiment is to
express and purify recombinant His tagged, Green Fluorescent Protein
(GFP). GFP is a protein native to a jellyfish, Aequorea <i>Victoria</i>and due to its remarkable fluorescence it has a vast amount of
applications in modern biochemistry such as in vivo labelling and
visualization of cells structures. Since the GFPs fluorescence disturbed
by unfavorable conditions can recover in the correct pH and temperature
it is incredibly useful in molecular biology and its ability to track
“real time” changes in cells with a “naked eye” captured the
attention of various researchers. (Wu et al., 2008) Another big
advantage of using GFP over other molecular trackers is that it does not
require any substrates or cofactors has a low toxicity towards examined
cells and does not alter the localization of its fusion partner. (Kumar
et al., 2016)</div>