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Lab #7: Transform E. coli with GFP Plasmid Using Electroporation and Heat Shock
1. Laboratory #7 pGlo Transformation and Purification of Green Fluorescent Protein
(GFP)
Dr. Belinda Román
Transformation is the introduction of genetic material from one specie into
different specie. In this case, we used a pGLO plasmid from a jellyfish to transform the
bacterium E. coli. Since GFP is a visual marker, it is used to study biological processes of
the central framework of biology. GFP locates genetic material, synthesis of proteins, and
helps to observe gene expression. We conducted electroporation and heat shock.
Electroporation uses an electrical shock to make cell membranes permeable to DNA. The
heat shock, integrates GFP in the genome. For this experiment, we used two positive
pGlo and two negative pGlo. The positive pGLO are with LB/amp and LB/amp/ ara.
From these two the only that fluoresced was the LM/amp/ara, because ara is the sugar
that regulates proteins. The other two dishes are the negative controls lacking ara;
therefore, no fluorescence was visible.
