Conor Wight

Mrs. Lyon
AP Biology
1/28/2016
pGLO Transformation write up
Introduction
In this experiment, plasmids, promoters, antibiotics and
antibiotic resistance were all applied in order to achieve the result of
transformation. A plasmid is a small, circular piece of DNA that
comprises of genes that control one or more traits that can have a
positive impact on a bacterias ability to survive. In this experiment
specifically, the pGLO plasmid was analyzed. pGLO encodes the gene
for both a GFP and a gene for resistance to antibiotic ampicillin. As the
name suggests, an antibiotic is used to inhibit the growth of and/or
destroy bacteria, which makes the benefit of the pGLO plasmid obvious
in the face of ampicillin. In addition to resisting antibiotics, pGLO
employs a gene regulation system in which a promoter, a sugar called
arabinose, is used to control the expression of the fluorescent protein
(GFP) in transformed cells. All of these factors contribute to the
transformation of the E. Coli bacteria. Transformation is the action of
inserting a gene into an organism in order to alter or create a
phenotype in a given organism. In this case, the E. Coli bacteria was
transformed in order to give it the ability to produce bioluminescence,
the same trait found in many jellyfish, among other organisms.
Petri Dish Physical Descriptions
Petri Dish
Colour
Growth
+pGLO
White, no
Large
LB/amp
glow

Pictures

+pGLO
LB/amp/ara

Yellow, glow

Large

-pGLO LB/amp Yellow, glow

Growth*

-pGLO LB

Lawn
growth

Yellow, no
glow

*No growth should have occurred as the ampicillin was accidentally

killed off, making conditions identical to the -pGLO LB plate
Transformation was successful. This is due to the fact that the
coloration of the bacteria became white whe n the plasmid and
ampicillin was added to the bacteria, while the bacteria in the second
petri dish exhibited similar coloration and the trait to glow under a UV
light as a result of the arabinose sugar. Both of these were the desired
phenotypes from the attempted, and successful, transformation.
This lab could be made more successful with more trials being
run, as well as the possibility of better backdrops and/or lighting in
order to truly see the desired colors, as each plate looked relatively the
same to the unprofessional eye. This would have also eliminated the
inevitable human error over the timing of the heat shocking of the
plasmid.
Organisms stand to gain from the ability to turn a gene on or off
in the face of varying conditions. One such example of this would be in
the realm of camouflage; the ability to turn a gene on in order to

match the coloration of an organisms surroundings is advantageous

for both predators and prey.
One potential application of transforming bacteria exists in
medicine, a field in which bacteria can be genetically altered to assist
humans against illness. If transformed with the correct genes, bacteria
can be applied within medicines and vaccinations in order to stop
diseases and viruses from afflicting humans and other organisms.