Chelsea Hawkins

BIOL 4915 1

4/23/15

Proposal: Can the Consumption of Genetically Modified Corn Be Linked to Infertility

Studying the Glyphosate Tolerant Form of the CP4 EPSPS Gene

Purpose and Objectives

The population of people affected by foods with genetic modification from glyphosate is
growing. The concept of this experiment is to link glyphosate to the infertility problems both
males and females experience. In the experiment performed in this lab, the objective was to
identify the food products as being genetically modified or not genetically modified. By
established this initial identification and by focusing on specific food products, the research will
be more concentrated on its specific topic. This proposal is to secure funding for research in the
biomedical science field studying the CP4 EPSPS Gene. By studying this enzyme, as it is linked
to glyphosate, this research has the potential to further investigate the subject of infertility.
Background
Maize is among the most common genetically modified plants. There are several ways to
genetically modify a plant, one being to spray it with an herbicide. Glyphosate is the main
ingredient to most herbicides. Farmers use these herbicides to save their crops from things such
as being overgrown by weeds. Even though this product was developed for progressive purposes,
it poses detrimental health effects to both the farmer and the consumer of the product. By using
glyphosate to spray their crops, farmers prolong their exposure to the toxins that are affecting
them and being brought back into their homes to affect their families. Glyphosate is modified
with unknown adjuvants and consequently introduce genes into the plant. Once the genetically
modified food is consumed, the effects associated with the specific gene introduced into the plant

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can either disturb the biological processes of the human body or the effects of the glyphosate can
be more severe than the gene itself.
Genetically modified plants exhibit some similarities. One of those similarities is their
promoter and terminator sequence. If a plant is genetically modified, it will show the presence of
the CaMV35S promoter, Cauliflower Mosaic Virus, and NOS terminator, Nopaline Synthase.
This promoter and terminator are signs that a plant has been genetically modified. These results
would be proven in the DNA Gel Electrophoresis. The bands from the DNA gel would confirm
the presence of these sequences. Confirming the presence of these sequences in the food
products means that the food contains a modified gene.
The gene studied in this experiment is known as CP4 EPSPS (5-enolpyruvulshikimate-3phosphate synthase). It is a strain derived from Agrobacterium tumefaciens. The function of this
gene is to decrease the binding affinity for glyphosate, which in turn increases the plants
tolerance to the glyphosate herbicide. The gene itself is said to have no effect in humans. The
EPSPS enzyme is part of the shikimate pathway that is involved in the production of aromatic
amino acids and other aromatic compounds in plants (ILSI Research Foundation, 2015). It is
believed that the shikimate pathway is not present in animals and therefore does not pose a
problem when consumed by humans. However, it is ordinarily overlooked that this pathway is
present in human gut bacteria (Feeney, 2014). Thus, meaning that this gene may indeed have an
effect upon human biological processes.
Experimental Design/Budget
Maize is the chosen plant to be studied for this experiment. Three food products derived
from this plant were tested for genetic modifications and plant specificity. These three products

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were frozen organic sweet corn, yellow corn meal, and yellow corn bread mix. These products
were chosen based upon their DNA reliability and their popularity in households. Frozen organic
sweet corn was chosen because of the movement for preference toward organic products.
Organic products are being favored as the approval of new genetically modified products is
increasing. Yellow corn meal and corn bread mix were chosen because of their many uses in
recipes for other foods.
This experiment began with the extraction of DNA from the three chosen food products.
To begin extracting DNA, the food was crushed in a mortar and pestle, mixed with water and
InstaGene Matrix buffer solution, heated, and then centrifuged. After being centrifuged, a small
amount of supernaut was taken from the screw tube and placed into two PCR tubes. The DNA is
placed in two different PCR tubes because two primers are used. The GM primer tests the sample
for genetic modifications. Specifically, the GM primer test for the CaMV35S promoter and NOS
terminator sequence. These sequences are common to genetically modified plants.
DNA was also extracted from a non-GM food and used as a control for GM negative
results. The GM positive control was a sample of ready-made DNA. Each sample was also
placed in two separate PCR tubes to be mixed with the GM and PS primer. After running the
PCR, the products were then placed into agarose gel wells to be run by DNA Gel
Electrophoresis. The results are to be found within the bands on the gel.
The estimated operating budget for the research conducted includes the following:

Mortar & Pestle: $4.00

Corn Food Products: $15.00
Lab Scale: $160.00
Pipets/Tips: $950.00
Centrifuge: $550.00

Chelsea Hawkins

BIOL 4915 1

4/23/15

GMO Investigator Kit: $160.00

PCR: $2,500.00
Agarose: $60.00
Flask: $20.00
TAE: $20.00
Gel Electrophoresis: $275.00
Fast Blast Stain: $24.00
ChemiDoc Imaging System: $20,000.00

The estimated total budget for this experiment is $24,738.00.

Potential Results
The results from this experiment are to be found in the agarose gel. DNA Electrophoresis
was run on the gel. The gel was then stained with Fast Blast Stain. After de-staining the gels, the
results could be read based on the bands within the gel. The ChemiDoc Imaging System was
used to prepare contrasted images of the gel. On the first gel, Test Food 1 ran with the GM
primer does not show any bands. However, when ran with the Plant Specific primer, one
moderate band shows. On the second gel, Test Food 2 shows a faint GM positive band. Test food
3 shows a band from being run with a Plant Specific primer. These results provide insight on the
nature of these corn products and whether their source was genetically modified or not.

Impact
The impact of this research would reach far into all communities. Infertility has always
posed a problem regardless of gender, race, or ethnicity. The world is filled with people who are
ready to have children and have been attempting to have children. After a while of trying to have

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children, they book an appointment to see a fertility specialist, who does all they can to give
them what they are trying to achieve. Here is something to think about: What if something you
were eating caused you to become infertile? This research would address this issue. Perhaps
glyphosate has entered into your system and disrupted your reproductive functions. Would there
be a way to detect if this was the cause? Most research articles focus solely on male infertility.
This research would address both male and female infertility. There are reports that establish a
basis for disrupted reproductive function in animals caused by pesticides. The validation of these
concepts is prominent and would be a step forward in preventing detrimental glyphosate health
effects.

Chelsea Hawkins

BIOL 4915 1

4/23/15

Table 1. PCR Cycling Conditions

Figure 1. Laboratory Procedures

References
Table
Table
2. DNA
3. DNA
Gel Electrophoresis
Gel Electrophoresis
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Pattern:
Pattern:
Gel #1
Gel #2
Busbee, D. C. (2007).

Alteration

of estrogen-regulated gene expression in human cells induced by the agricultural and

horticultural herbicide glyphosate. Human & Experimental Toxicology, 747-752.
Retrieved from

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http://web.a.ebscohost.com.ezproxy.ololcollege.edu:2048/ehost/pdfviewer/pdfviewer?
sid=45af106b-d034-499d-a214-51636e76eb2e%40sessionmgr4004&vid=1&hid=4204
Center for Environmental Risk Assessment. (2010). A Review of the Environmental Safety of the
CP4 EPSPS Protein. Washington, DC: ILSI Research Foundation. Retrieved from
http://cera-gmc.org/docs/cera_publications/pub_01_2010.pdf
Feeney, D. F. (2014, April). How Roundup (AKA. Glyphosate) Affects Your Health. Nutritional
Perspectives: Journal of the Council on Nutrition, 33. Retrieved from
http://web.a.ebscohost.com.ezproxy.ololcollege.edu:2048/ehost/pdfviewer/pdfviewer?
sid=3315ae23-6fc8-4ff7-b11a-45fd66cd5572%40sessionmgr4001&vid=1&hid=4204
ILSI Research Foundation. (2015, February 27). GM Crop Database. Retrieved from Center for
Environmental Risk Assessment: http://ceragmc.org/GmCropDatabaseEvent/MON89788