Objective:

The objective of my research is to screen potato-related food products sold on the market
for Cry3a gene. Four potato products will be used to find this gene: fresh potatoes, potato chips,
boxed mashed potatoes, and frozen French fries. The potato products are attained from Walmart,
Whole Foods, Trader Joes, and Southside Produce.
Hypothesis:
Cry3A gene is present in all food products, purchased from local stores, whether they are
labeled organic or not.
Background:
One of the many reasons why certain crops are being genetically modified is to protect
them from pests. One of the potato crops biggest pests is the Leptinotarsa decemlineata, or also
known as the Colorado potato beetles. Adult female Colorado potato beetles lay their eggs on
potato leafs and both the larvae and adult Colorado potato beetles feed on their nested potato
leafs (Jacques & Fasulo, 2015). Cry3A gene in Bacillus thuringiensis (Bt) is inserted to some of
the seeds of some potato crops in order to release a toxin that is very harmful to Colorado potato
beetles. My mentor, Dr. Gupta, provided an easy-to-understand visual (attached on reference
page) on how a gene can be harvested and transferred into plants. According to Mi et al., (2015)
Once ingested by susceptible insects, Cry proteins would have been proteolytically activated by
gut proteases and the active toxin would have bound to receptors located on the brush border
midgut epithelium to exert their toxicity, and finally broken down the gut epithelium of insects,
leading to their death.

Since Bt toxin is one of the most common toxins that is released in genetically modified
plants to fight off pests, researchers have been studying its side-effects on both human health and
environmental risks. In a study done by Ostr, Mal, & Pfohl-Leszkowicz (2014), 34 different
genes with unique functions are harvested from Bt is inserted to maize crops. Using several
varying analytical methods, they find that maize crops that have been genetically modified (have
Bt genes inserted to them) contain less aflatoxins (toxins which are linked to liver cancer) than
maize crops that have not been genetically modified. Mesnage et al. (2013) experimented on
embryonic kidney cell line interactions with Cry1Ab and Cey1Ac Bt toxins and their combined
actions with Roundup (a pesticide). Cry1Ab toxin alone causes cell death, while Cry1Ac toxin
has no effect. They also found that the combination of Roundup and both gene toxins delay
activation of apoptosis. As far as the environment goes, Tharafdar, Rathore, & Shiva (2012)
noted in their study that there is no difference in soil characteristics in genetically modified
cotton crops vs. non-genetically modified cotton crops. However, both enzyme and microbial
activities are greatly reduced in the genetically modified crops soil as opposed to the nongenetically modified crops.
Experimental Design and Budget:
The target DNA sequence of the target gene can be found through the GenBank. DNA
from chosen potato-based food products will have to be extracted first. A small piece of the food
product to be tested is placed inside a mortar with some water added to it. The water and food
product is then grinded into slush. Once that is done, the mixture is heated and centrifuged. The
supernatant resulted after centrifugation is the DNA extract. PCR is then used to amplify the
DNA. During PCR, the DNA gets denatured at around 94C, annealed at 59C, and extended at
72C. The promoter being used for this study is Cauliflower Mosaic Virus (CaMV) 35s and the

terminator used is Nopaline Synthase terminator, or also known as NOS terminator. The CaMV
promoter is used instead of any other promoter because it is the most powerful and is not greatly
influenced by environmental conditions or tissue types. The promoter is about 400 base pairs in
length. NOS terminator is used for this experiment because it is the most common one to use for
screening genetically modified foods. The sample received from PCR is then transferred on to
electrophoresis gel to be read.
Through field research, I find that the prices of the food products vary from store to store.
Fresh potatoes sell for 59.4c/lb. in Walmart, $1.99/lb/ in Whole Foods (potatoes from this
specific store is labeled organic), 69c/lb. in Southside Produce. Our local Trader Joes do not
carry fresh potatoes. A Walmart brand potato chips go for $2.00 per 16oz. bag (12.5c/oz.), an
organic-labeled Whole Foods brand potato chips go for $2.87 per 5oz. bag (57.4c/oz.), and a
Trader Joes brand potato chips (also labeled organic) cost $2.29 per 10oz. bag (22.9c/oz). Boxed
mashed potatoes cost $2.68 per 26.7oz. in Walmart (10c/oz) and organic-labeled boxed mashed
potatoes from Whole Foods and Trader Joes cost $1.99 per 8oz. box (24.9c/oz.) and $2.29 per
28oz. (8.2c/oz.) respectively. A Walmart bag of frozen French fries cost $4.48 per 80oz. bag
(5.6c/oz.), $2.99 per 32oz. bag (9.3c/oz.) at Whole Foods, and $1.99 per 24oz. bag (8.3c/oz.) at
Trader Joes. A website for a company called Thomas Scientific sells a CaMV promoter and
NOS terminator kit complete all the components used for gel electrophoresis for $939.00 and
without gel electrophoresis components for $741.00.
Impact:
Many of the food products, from fresh ingredients to bagged, that are sold in stores today
contain genetically modified organisms. If a food product does not contain genetically modified

organisms (GMOs), why not? The term GMO seems to have a rather negative connotation,
mostly because not many people are educated on the basic of how GMOs even work. After being
exposed to people who heavily promote non-GMO products, one of the main reasons why many
people avoid GMOs is the fact that not a lot is known about the effects of consuming GMOs.
Genetically modifying our food is a relatively new technology and there are not many researches
done to study the long-term effects of consuming genetically modified organisms. In the long
run, I hope that we, as human beings, find all the side effects of every toxin released by the genes
we insert to our food. With the knowledge, I then hope that we can sedate their harmful effects so
that genetically modifying our food is common sense. The benefits of GMOs greatly outweigh
the risks. With the advancement of medicine and antibiotics, the human population does not
seem to stop growing anytime soon which means that we need more and more space. Although
we can expand vertically (i.e. apartment complexes), growing our crops vertically is not feasible.
The best viable option for us is to genetically engineer our crops so that we can get the nutrition
that would sustain our never-ending population growth.
Potential Result:
A lot can happen during the experiment. Although I have learned the theory of how each
machine works, this experiment will be the first time I ever use them and therefore mistakes are
bound to happen. Since our resources are limited and time is of the essence, I would not be able
to repeat and fix my mistakes. However if everything goes smoothly, four possible results can be
achieved. Screened products that are labeled organic will show no presence of the Cry3A gene,
while the ones that are not labeled organic will show presence of Cry3A gene. A second
possible result would be the products labeled organic contains Cry3A gene and unlabeled

products do not. Third possible result is that are both labeled and unlabeled as organic do not
contain the Cry3A gene. Lastly, both labeled and unlabeled products contain the Cry3A gene.

References
Jacques, R. L., & Fasulo, T. R. (2015, November). Colorado potato beetle - Leptinotarsa
decemlineata (Say). Retrieved from
http://entomology.ifas.ufl.edu/creatures/veg/leaf/potato_beetles.htm
Mesnage, R., Clair, E., Gress, S., Then, C., Szkcs, A., & Sralini, G. (2013). Cytotoxicity on
human cells of Cry1Ab and Cry1Ac Bt insecticidal toxins alone or with a glyphosatebased herbicide. Journal Of Applied Toxicology, 33(7), 695-699. doi:10.1002/jat.2712
Mi, X., Ji, X., Yang, J., Liang, L., Si, H., Wu, J., & ... Wang, D. (2015). Molecular biology and
genetics/Biologie et gntique molculaires: Transgenic potato plants expressing cry3A
gene confer resistance to Colorado potato beetle. Comptes Rendus - Biologies, 338443450. doi:10.1016/j.crvi.2015.04.005
Ostr, V., Mal, F., & Pfohl-Leszkowicz, A. (2015). Comparative data concerning aflatoxin
contents in Bt maize and non-Bt isogenic maize in relation to human and animal health a review. Acta Veterinaria Brno, 84(1), 47-53. doi:10.2754/avb201585010047
Tarafdar, J. C., Rathore, I., & Shiva, V. (2012). Effect of Bt-Transgenic Cotton on Soil Biological
Health. Applied Biological Research, (1), 15.