Genetic Transformation Of E. Coli

1166 WordsNov 30, 20155 Pages
GENETIC TRANSFORMATION OF E. COLI WITH pGLO AS A VECTOR USING THE HEAT SHOCK METHOD Katelyn Brown, L07 Introduction: Escherichia coli is a small, usually harmless bacterium that commonly resides in human intestines. First discovered by Theodor Escherich in 1885, it has become a widely used organism in the science world. It is especially efficient in scientific experiments because it grows easily and quickly in a stimulating environment, is usually harmless, and is very easy to obtain (Berg 2004). Escherichia coli (E. coli) is very susceptible to genetic transformation, which is what this experiment focuses primarily on. (Hanahan 1983). This experiment was performed in order to see if E. coli can be transformed by adding an ampicillin resistant plasmid and using heat shock to increase uptake. Heat shock is a method in which a cell is placed in an environment of elevated temperature. This can restructure the cell wall and make it more permeable, and it can also amplify gene expression (Verghese et al. 2012). Thus, heat shock is used in this experiment in order to attempt to amplify an induced gene expression of fluorescence in E. coli. A plasmid called pGLO is used as the vector in this experiment. pGLO contains two genes that are vital for this experiment; one that provides resistance to ampicillin and the green fluorescent protein (GFP) gene. The GFP gene is derived from the luminescent jellyfish Aequorea victoria. Because E. coli is a bacteria, it cannot survive when
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