Screening of Food Products Containing CP4 EPSPS Gene

Chelsea Hawkins, BS Student

Our Lady of the Lake College, Baton Rouge, LA
Abstract

Methods

The population of people affected by genetically modified (GM) foods from

adjuvants which introduces genes into the food we eat. The consequences of the
consumption of GM food can lead to the introduced gene disturbing the biological
processes of the human body or the effects of the glyphosate can be more severe
than the gene itself. The proposed concept in this experiment was to find an

Add 5000L of water per gram of food

Grind mixture until smooth slurry forms

Add additional 5000L of water per gram of

by pesticides.

Introduction

TF1

DNA

20

TF1
20

20

PSP

-----

1.45

-----

-----

-----

1.45

-----

1.45

+ 500L of InstaGene Matrix buffer

MM

19.55 19.55

21

16

21

16

19.55 19.55 19.55 19.55

Heat 5-6 minutes at 95

Total

45

40

45

40

Centrifuge at 6000 rpm for 5 minutes

Take 50L of supernatant and place 20L in

Take 250L of slurry and add to screw tube

Glyphosate linked to male infertility in low doses, more specifically Roundup

Health Effects: Birth Defects, Cardiovascular Disease, Cancer, and Alteration

41

41

Function Temp.

Duration

Cycles

Denaturation

Denature 94C

2 minutes

Denature 94 C

1 minute

Amplification

Mosaic Virus, and NOS terminator, Nopaline Synthase

tolerance to herbicides/pesticides

59 C

1 minute

Extend

72 C

2 minutes

Extension

Extend

72 C 10 minutes

Hold

Hold

4C

Pour mixture into gel tray (Sit 15 20 minutes)

to play a prominent role in the research of associated health effects. The impact of

Place gel in electrophoresis tray.

this research would reach far into all communities. Infertility has always posed a

Fill tray with 1x TAE buffer.

problem regardless of gender, race, or ethnicity. This research would address the

Mix 10L of orange loading dye with each tube

question, Is what you are eating effecting your fertility? This experiment poses

Pipet 20L of MWR and each PCR product into respective wells

the question for future research that would address both male and female

Run electrophoresis at 100V for 30 minutes

infertility. The validation of these concepts is prominent and would be a step

Stain gel with Fast Blast, de-stain with warm water

forward in preventing detrimental glyphosate health effects.

Use ChemiDoc to take image of gel

References
glyphosate. Human & Experimental Toxicology, 747-752. Retrieved from
http://web.a.ebscohost.com.ezproxy.ololcollege.edu:2048/ehost/pdfviewer/pd
fviewer?sid=45af106b-d034-499d-a214-51636e76eb2e
%40sessionmgr4004&vid=1&hid=4204

Results

2. Center for Environmental Risk Assessment. (2010). A Review of the

TF1 displays no band (negative) when tested with Genetically

Environmental Safety of the CP4 EPSPS Protein. Washington, DC: ILSI

Modified Primer.

Research Foundation. Retrieved from http://cera-

TF1 displays a band (positive) when tested with Plant Specific

gmc.org/docs/cera_publications/pub_01_2010.pdf
3. Feeney, D. F. (2014, April). How Roundup (AKA. Glyphosate) Affects Your
Health. Nutritional Perspectives: Journal of the Council on Nutrition, 33.

TF2 displays a faint band (positive) when tested with Genetically

http://web.a.ebscohost.com.ezproxy.ololcollege.edu:2048/ehost/pdfviewer/pd
1000bp

TF2 displays no band (negative) when tested with Plant Specific

700bp

Primer.

500bp

Test food is not plant specific, but does exhibit genetic modifications.

This pathway is present in human gut bacteria; therefore it can attribute to

GMP: Genetically Modified Primer

40

Modified Primer.

1000bp

other aromatic compounds in plants

MWR: Molecular Weight Ruler

households. The knowledge of which foods are genetically modified will continue

Retrieved from

500bp

GM(O): Genetically Modified (Organism)

Indefinite

DNA
Electrophoresis
Heat mixture
and allow to cool, but not solidify

1. Busbee, D. C. (2007). Alteration of estrogen-regulated gene expression in

Test food is plant specific, but does not exhibit genetic modifications

Shikimate Pathway: Involved in the production of aromatic amino acids and

Legend:

41

Primer.

700bp

the associated health effects

41

The foods in this experiment were chosen based upon their commonality to

human cells induced by the agricultural and horticultural herbicide

Anneal

glyphosate

Function: Promotes a decrease in binding affinity, which increases a plants

41

Step

EPSPS gene is present in plants, but modified to CP4 EPSPS to tolerate

41

PCR Cycling Conditions

to Endocrine, Neurological, and Reproductive functions

Foods: Alfalfa, Cotton, Maize, Potato, Soybean, Sugar Beet, and Wheat

20
-----

Repeat with each food.

20
1.45

CP4 EPSPS: 5-enolpyruvulshikimate-3-phosphate synthase enzyme

20
-----

Almost all GMOs are modified for herbicide/pesticide tolerance

20
1.45

two PCR tubes.

Terminator
(NOS)

20
-----

CP4 EPSPS

20

GM+ GM+ GM - GM -

Rinse M&P with 10% bleach and water

Promoter
(CaMV35S)

TF3

-----

Most common GMOs include: corn, soy, canola, sugar beets, cotton, and

A GM plant will show the presence of the CaMV35S promoter, Cauliflower

20

TF3

TF2

-----

papaya

TF2

Mix 70mL of 1x TAE buffer with 2.1g agarose

1.45

reports establishing a basis for disrupted reproductive function in animals caused

(in L)

GMP

food

association between the consumption of GM foods and infertility, as there are

PCR Preparations

DNA Extraction

the pesticide glyphosate is growing. Glyphosate is modified with unknown

Discussion

100bp

TF3 displays no band (negative) when tested with Genetically

Modified Primer.

TF3 displays a band (positive) when tested with Plant Specific

Primer.

Test food is plant specific, but does not exhibit genetic modifications.

45fd66cd5572%40sessionmgr4001&vid=1&hid=4204
4. ILSI Research Foundation. (2015, February 27). GM Crop Database.
Retrieved from Center for Environmental Risk Assessment: http://cera-

200bp

200bp

fviewer?sid=3315ae23-6fc8-4ff7-b11a-

gmc.org/GmCropDatabaseEvent/MON89788

100bp
Test Food 1 (TF1): Cornbread Mix
Test Food 2 (TF2): Yellow Corn Meal
Test Food 3 (TF3): Frozen Organic Sweet Corn