Transformation of E.coli HB 101 with PGLO

By Hewan WamiBridges 2014

Prof. Sara F.Feinman Prof. Heather Kundzicz

June 13, 2014

My Mentor

William E. Robinson, PhD

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).

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

Transformation of E.coli with PGLO plasmid

The pGlo plasmid also contains the Green Fluorescent Protein (GFP) gene. GFP gene, which is derived from a jellyfish called Aequorea Victoria,

◦ 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.

Green Fluorescent Protein

Thank you