Hallie Ryan

Mrs. Peterson
AP Biology
February 3, 2016

pGLO Transformations Lab

Introduction: Bacteria contain genes for traits that may be beneficial to the
bacterial survival. Bacteria can transfer plasmids back and forth, which allow them to
share these genes. In this lab, the pGLO is resistant to the ampicillin. Therefore, the
plasmid gives resistance to the antibiotic on the plates that have amp. Antibiotic
resistance happens when an antibiotic loses its ability to control/stop bacterial growth.
The transformation in this lab describes how effective the transfer the plasmid into
bacteria was. Gene transformation involves the insertion of a gene into an organism,
which is supposed to change one or more of the organisms traits. In the lab, the bacteria
were transformed with a gene that coded for GFP. The gene for GFP can be turned on in
the transformed cells by adding ara. The promoter either will prevent or allow
transcription of GFP. In the absence of ara, the araC regulatory protein binds to the
promoter which prevents transcription, but the in presence of ara, the regulatory protein
(araC) can bind to the ara, which changes the protein. This makes many GFP proteins,
meaning it glows. In other words, the ara starts the transcription of the genes by
promoting the binding of the RNA polymerase. In the pGLO plasmid, some of the genes
that help with the breakdown of ara were replaced with GFP. When the bacteria have
been transformed with the plasmid are grown in the presence of ara, the GFP is switched
on and the bacteria will glow under the UV light.
Data Collection and Analysis of the +pGLO cells with the non-transformed E.
Coli
Pictures

+pGLO LB/amp

General
Growth

Color (with
and without
UV light)
A lot of growth Under UV
because has the light there is
resistance gene no glow
because no ara

+pGLO
LB/amp/ara

Amp +
restriction
they cancel so
it shows a lot
of growth

Has resistance
gene and ara,
meaning it
glows under
the UV light

-pGLO LB/amp

Since there is
No glow under
no plasmid and UV light
the plasmid
does not have a
resistance
gene, it means
there is no
growth.

-pGLO LB

Since theres
No glow under
no amp, theres the UV light
nothing
restricting it.
This means
there is
growth.

Conclusion
The organism can adapt to different conditions and will not have to waste energy
on producing unnecessary proteins. Also organisms don't always want the expression of
every gene, so in certain condition this will be beneficial to them because they will not
have to express every gene. To improve the experiment and get better results next time,
the incubation time could have been more exact. If it wasnt exact the first time doing this
experiment then there was room for error. Also maybe microbes/outside bacteria could
have gotten into the plates in they were open for too long which could have affected the
results of the experiment as well. There are many potential applications of transforming
bacteria. Firstly genetic engineering allows the transformation of bacteria with the gene
coding for a useful protein. Then, growing the transformed organism can produce large
quantities of the protein. Also by using gene-splicing scientists can manufacture mass
amounts of insulin by using the cells of E. coli bacteria. It can be determined if the E. coli
has been genetically transformed by inserting a new piece of DNA that will make the
bacteria glow. The E. Coli wouldnt have had a transformation if the pGLO plasmid
hadnt been added. From this lab it was shown that the bacteria was amp resistant since
the bacteria kept growing and transforming. The E. coli on the LB/amp/ara became
florescent under the UV light, meaning the transformation had worked. It grew with the
amp in the agar showed that the plasmid produced an enzyme that works against the
antibiotic amp.