pGLO Transformation Lab

www.welc.cam.ac.uk/ ~pineslab/Projects/gfp.htmRenilla reniformis

www.wileywiggins.com/ 2003_05_01_blog2.html

www.worldnetcams.com/sealife/cerianthus.jpg

www.afae.it/pages/galleria%20immagini/Invertebrati%20marini/trachy.htm

Jellyfish

CopepodsOceanexplorer.noaas.gov/explorations/05deescope/background/fluorescence

AmphipodsOceanexplorer.noaas.gov/explorations/05deescope/background/fluorescence

Corals

Many organisms can glow! Many organisms can glow!

Crystal Jelly has a gene called GFP that makes it glow!

when something glows, it is fluorescent

Transgenic MiceTransgenic = something that has genes from different organisms (individuals)

To remember...trans = movegenic = about genes

So transgenic = genes moved from one organism to another

E. coli = bacteria

bacteria = very small organisms that have DNA but no nucleus

They were some of the first life on earth!

In this lab, we will make bacteria glow !!

Why should we do this? TO DO BIOTECHNOLOGY!

DNA RNA Protein Trait

GFP Gene

• found in jellyfish

• put into bacteria

Green Fluorescent Protein

GLOWING CELLS

Bacteria now make protein that glows…

Bacterial chromosomeTransformation is

when the DNA from one organism is combined with the DNA from another organism.

Bacterial chromosome

How will we do this? Transformation.

Plasmid with recombinant DNA!

STARTING MATERIALS

Bacterial chromosome

E. coli cells

• sensitive to antibiotics

• can’t glow

• can be transformed

END RESULT

AmpR

Ara

GFP

Bacteria with Recombinant DNA…

… that can GLOW!

GROW ON AN AGAR PLATE

Genes on the Plasmid

AmpR

Ara

GFP

pGLO plasmid

makes all transformed bacteria resistant to ampicillin

Makes the bacteria glow

Helps the bacteria make GFP when it eats the sugar arabinose