Annie Wallace

pGLO Transformation Lab

Introduction:
This experiment involved many factors, which caused varying
results in the four different petri dishes. A plasmid is DNA that can
replicate independently and is often is the avenue for which bacteria
gains resistance to an antibiotic. To make recombinant DNA you must
cut two different plasmids with the same restriction enzyme. In this
lab, the plasmid is pGLO, which contains the genes for Green
Fluorescent Protein (GFP). The promoter controls initiation of
transcription, which creates the RNA. This gene is the gene of interest
and has a code for a gene for resistance to the antibiotic ampicillin.
The plasmid was pGLO, which was present in two of the four petri
dishes. E. coli can be transformed to make the GFP protein and express
the gene, which would cause the plasmids to glow when exposed to UV
light. In certain agar plates with ampicillin, there will be no bacterial
growth because it is an antibiotic that is resistant to the plasmid. In the
experiment, the GFP gene can be switched on or off in transformed
cells by adding sugar arabinose to the nutrient medium. In the
experiment, agar plates with +pGLO LB/amp will have the most
success because they have the resistance gene, therefore allowing the
bacteria to grow.
Results:
Transformation of Bacteria on Petri Dishes with Different Factors
Petri Dish:
Results:

+pGLO LB/amp

+pGLO
LB/amp/ara

-pGLO LB/amp

-pGLO LB

Observation
s:

A lot of bacterial
growth, more
colonies reproduce
Bacteria doesnt
glow because no
arabinose

A lot of
reproduction,
colonies
everywhere
Bacteria glows
due to presence
of arabinose

No growth of any
bacteria, no
pGLO (resistance
gene)

A lot of growth,
colonies
everywhere
(lawn)
Does not glow

Conclusion:
In this lab, transformation was successful. It is clear that it
occurred because there would be no bacterial colonies on the plate
with the pGLO and ampicillin if transformation had not occurred. The
plasmids on the this plate would need to have recombinant DNA on it
in order to survive in the presence of the antibiotic, whose purpose is
to stop continuous growth. The two plates that contained the plasmid
both had bacterial resistance to the antibiotic. Additionally, the plates
with the plasmid were fluorescent in the end. To improve the
experiment next time, it should be ensured that all the tools are sterile,
E. coli is treated correctly and correct amounts are used of each
material to provide optimal success. The advantage of being able to
turn on and off a gene can contribute to maximum survival potential.
Animals can change a particular gene depending on the conditions
they are enduring. For example, if an animal was able to become
invisible at certain times, it could escape predators very easily, and
they would thrive. Transforming bacteria is very common in food.
Farmers often insert bacteria into their crops in order to modify the
food to be able to grow rapidly and survive during certain weather
conditions. This is very beneficial for the farmers because they can
make a very large profit by genetically modifying the food and
inserting bacteria into it.