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Transformation of E.coli HB 101 with PGLO. By Hewan Wami Bridges 2014 Prof. Sara F.Feinman Prof. Heather Kundzicz June 13, 2014. My Mentor. Professor of Environmental and Aquatic Toxicology, School for the Environment, College of Science and Mathematics
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Transformation of E.coli HB 101 with PGLO By HewanWami Bridges 2014 Prof. Sara F.Feinman Prof. Heather Kundzicz June 13, 2014
My Mentor • Professor of Environmental and Aquatic Toxicology, School for the Environment, College of Science and Mathematics • His research addresses functional mechanisms in aquatic toxicology, particularly those processes involved in metal uptake, depuration, sequestration and internal transport. • This work focuses on two groups of marine organisms – • bivalve molluscs (mussels and clams) and • tunicates (ascidians). William E. Robinson, PhD
Transformation of E.coliwith PGLO plasmid • Transformation is a process in which cells take up foreign DNA from their environment. • Transformation can occur in either prokaryotic or eukaryotic cells • E. coli is sensitive to an antibiotic called ampicillin. • Ampicillin interferes with the formation of bacterial cell walls and thus kills newly divided cells that must form new cell walls. • The pGlo plasmid contains an ampicillin resistance gene. • Ampicillin resistance gene encodes an enzyme, β lactimase
Green Fluorescent Protein • The pGlo plasmid also contains the Green Fluorescent Protein (GFP) gene. • GFP gene, which is derived from a jellyfish called AequoreaVictoria, • encodes a fluorescent protein that glows green when exposed to ultraviolet light. • The pGlo plasmid also contains the arabinose promoter. • A promoter is a short region of DNA that regulates the expression of a gene. • The arabinose promoter controls the expression of the GFP gene in the pGlo plasmid.