Literature Review : Green Fluorescent Protein And Histidine Tagged Proteins

1537 Words Jan 27th, 2015 null Page
Literature Review: Green Fluorescent Protein and Histidine Tagged Proteins Histidine and green fluorescent protein (GFP) tagging are important in molecular biology because they allow for purification, tracking, and quantification of target proteins (Ferrari et al., 2004; Cho et al., 2011; Deponte, 2012). Histidine tagging provides a method for isolating and increasing the amount of target protein recovered from a biological organism or mixed sample of proteins (Masek et al., 2011; Singh and Jain, 2013). GFP tagging allows target proteins to be tracked, quantified, and observed within a living organism (Deponte, 2012). Histidine tagging works by attaching a specific number of histidines to the carboxyl or amino terminus of a target protein sequence, promoting the extraction of the protein through molecular modification and chromatography techniques, mainly, immobilized metal ion affinity chromatography (IMAC) (Myscofski et al., 2000; Bornhorst and Falke, 2010; Singh and Jain, 2013). This can be done by attaching a DNA sequence encoding histidines to an expression vector and then inoculating the target protein in the mixture, allowing the protein to ligate to the histidine sequence and the staggered end in the expression vector (Myscofski et al., 2000). Then, when the vector is replicated and translated, the target protein is produced with a histidine tag and can be cut by restriction enzymes to isolate the target protein (Myscofski et al., 2000). For optimization of the…

Related Documents