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SamanthaWargo

050316
GeneticsLabBL213

AnalyzingifCornMuffinMixisGeneticallyModified

Introduction

Thisstudyanalyzesfoodtodetermineifitisgeneticallymodified.Geneticallymodified

organismsareorganismsthathaveDNAthathasbeenmodifiedinanunnaturalway,by

introducingnewgenes(WorldHealthOrganization,2016).Thefirstmodifiedfooditem,

tomatoes,wereapprovedtobesoldin1994(Laskos,2013).Somecommongeneticallymodified

cropsaremaize,soybean,cotton,andcanola.In2014,therewereatotalof12cropsthatwere

beingmodifiedandcommerciallyused(Bradshaw,2016).Aswithmostadvancesinscience,

therearemanyprosandconsthatgoalongwithgeneticallymodifiedorganisms.

Someoftheprosofusinggeneticallymodifiedfoodarelesspesticideuse,morenutrition,

anditsbetterfortheenvironment.Pesticidesareusedonplantstokillbugsthatwilltrytoeat

them.Pesticideusecanbeharmfultopeopleanddangeroustotheirhealth.Pesticidescanalso

pollutetheenvironmentordamagethesoil.Withgeneticallymodifiedfood,itcanbealteredso

thatpesticidesarenotneededtobeapplied(Havahart,n.d.).Forexample,Bacillusthuringiensis

canbeinsertedintothegenomeofcorn.Atoxicproteinwillthenbeproducedthatwillkillcorn

borersiftheytrytofeedonit.Bacillusthuringiensiscanbeinsertedintoothercropstoo,suchas

cotton(Qaim&Kouser,2013).Anotherprotogeneticallymodifiedfoodisnutritionvalue.

Somethirdworldcountriesareinneedoffoodandvaccines.Thecountriesaretoopoororthe

conditionsareunfavorablefornutritiousandplentifulfood.Foodcanbemodifiedsothat

caloriesornutritionvalueofcertainfoodswillgoup.Also,scientistscouldpotentiallyadd

vaccinestofood,whichwillbecheaperandeasiertodistributetothethirdworldcountries.

Finally,geneticallymodifiedfoodcanbebetterfortheenvironment.Notasmanyharsh

chemicalswillbeneededtouseonthecropsthusnotasmanyharshchemicalswouldbegoing

intotheenvironment.

Alongwithpros,therearealsoconstogeneticallymodifiedfood.Problemsthatcan

occurare:superweedsandsuperbugs,disruptiontotheecosystem,andantibioticresistance.

Superweedscouldoccurfromcrosspollinationbetweentheweedsandthegeneticallymodified

food.Superbugscouldbeproducedifthebugsbecomeresistanttothecompoundsinthecrops

thataremeanttokillthem.Alongwiththis,itcouldleadtodisruptionoftheecosystem.Some

believethatthisisalteringnaturewhichisdisruptingthenaturalenvironment.Alteringone

componentcanleadtoundesiredeffectsinothercomponentsfartherdowntheline.Anothercon

isthatantibioticresistancemaybecomeanissue(Mahgoub,2015).Antibioticresistantgenesare

usedasmarkersinthemodifiedorganisms,sothereisapossibilitythattheantibioticresistance

canthenhappeninotherplacesthanthecrops,inhumansforinstance(FoodandAgriculture

OrganizationoftheUnitedNations,2003).

Withgeneticallymodifiedorganisms,thereisaspecificterminatorand/orpromoterthat

ispresent85%ofthetime.Thecommonpromoteriscauliflowermosaicvirus,CAMV35S.This

geneisonthefiveprimeendandwillactivatetranscriptioninanyplant.Thecommonterminator

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isnopalinesynthase.Thisgeneisonthethreeprimeendandwillindicatewheretranscription

shouldend.

Someoftherisksassociatedwithgeneticallymodifiedcropsareunknownbecausethis

processisstillrelativelynew.Istheregoingtobelongtermeffectsonpeopleshealthorthe

environmentfromrepeatedexposure?Itshardtotellbecausealthoughtheyarenotseenyet,

unforeseenconsequencescouldstillshowup(Laskos,2013).Justbecausetherearenotany

negativeeffectsthatdoesnotmeantheyarenotstillthere.Scientistswillhavetotakethecases

astheycome,astheyarealreadytryingtocutofftheproblemsbeforetheyhappen(GreenFacts,

n.d.).

TheexperimentalobjectivesofthisexperimentincludeisolatingandamplifyingDNA

fromplantmaterialthroughpolymerasechainreaction(PCR)andgelelectrophoresis.Thepoint

oftheexperimentistotestafoodproductandconcludeifitisgeneticallymodifiedornot.Along

withthis,wecantesttheunknowntoseehowitisaffected.

Twomainprocedureswereusedinthisexperiment:PCRandgelelectrophoresis.PCR

worksbyfindingaDNAsequencethatisspecifictotheprimerandthenitamplifiesthis

sequence.ForPCR,thefoodproductsweregrindedupandmixedwithwaterandinstagene.The

instageneincludesdetergenttobreakthemembranes,bufferwithRNases,proteases,andDNase

inhibitor,andsomethingtobreaktheplantcellwalls.Theseslurrieswerethenmixedwiththe

PCRmastermixandranthroughthethermocycler,whichwaswheretheDNAwasamplified.

ThePCRmastermixincludednucleotides,theprimer,taqman,buffer,andmagnesium.Gel

electrophoresisuseselectricalcurrenttoseparateproteinproductbyweight.DNAisnegatively

chargedsoitmigratestowardsthepositivecharge.Thebiggerproductsmigrateslowerthanthe

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smallerproductsdo.Forthegelelectrophoresis,3%agarosegelwaspreparedwithgelreddye.

Thethreesamples,anegativecontrol,theunknownsample,andapositivecontrol,wereloaded

andran.

IhypothesizedthatcornmuffinmixwouldshowproteinproductfortheCAMV35S

and/ornopalinesynthasewhentheDNAwasamplifiedandanalyzedbecauseitisgenetically

modified.

Results

Inwelloneabandisshownat500bp.Inwelltwotherearenobandsshownexceptfora

bandinthedimerregion.Inwellthreethereisabandat500bp.Inwellfourthereisabandat

200bpandabandinthedimerregion.Inwellfivethereisabandat500bp.Inwellsixthereisa

bandat200bpandabandinthedimerregion.

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Figure1.AgaroseGelqualitativeanalysisindicatinggeneticallymodifiedfoodand

nongeneticallymodifiedfood.Wellsoneandtwoindicatethenegativecontrol,

nongeneticallymodifiedoats.Wellsthreeandfourindicatethecornmuffinmix.Wells

fiveandsixindicatethepositivecontrol.Theplantprimerwasusedinwellsone,three,

andfive.Thegeneticallymodifiedorganismprimerwasusedinwellstwo,four,andsix.

Gelreddyewasusedsothatthebandsarevisible.

Discussion

Thehypothesiswasacceptedbasedontheresultsachievedafterthegelelectrophoresis.

ThegelshowninFigure1indicatesthatthecornmuffinmixisgeneticallymodified.

Withgeneticallymodifiedfood,therewillbeabandsaround200bp.CAMV35Swill

appearat203bpandnopalinesynthasewillappearat223bp.AgaroseGelisnotidealto

indicateasmalldistancelikethis,butinourcasewedidnotcarewhichofthetwospecifically

showedup.Ourexperimentwasjusttoshowthattherewasabandinthatarea.At500bp,bands

willappearthatindicateaplantprimer.Theplantprimeristheretoindicatethatthetestis

workingcorrectly.Theplantprimerisoneforachloroplastgeneandwillshowupinallthree

plantsbecauseitisuniversalinallplantcells.

Asshowninfigure1,wellsoneandtwoweretheoats.Inwellone,thebandthat

appearedat500bpwastheplantprimer.Inwelltwo,abanddidnotappearat200bpsothat

meansthefoodwasnotmodified.Thiswasournegativecontrolforthestudy.Wellsthreeand

fourwerethecornbreadmuffinmix.Wellthreehasabandat500bpsothisshowsthatthetest

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wasworkingcorrectlyandthecornbreadmixisaplantproduct.Forwellfour,afaintbandcan

beseenat200bp.ThismeansthateitherCAMV35Sornopalinesynthaseispresentinthe

DNA,sotheplantisinfactgeneticallymodified.Wells5and6werethepositivecontrol,DNA

fromaknowngeneticallymodifiedfood.Wellfiveshowsabandat500bptoindicatethatthe

testisworkinganditisaplant.Wellsixhasabandat200bpsoitisgeneticallymodified.

Lowerthan200bpthereareseveralbandsshown.Thesebandsareindicativeofdimers

forming.Dimerscanformiftheprimerforthegeneticallymodifiedfoodreactswithitself.The

bandsappearlowerbecausethedimersareadifferentweightthanthemonomers.

Maizeisoneofthemostcommongeneticallymodifiedcropssoitmakessensethatcorn

muffinmixwouldgiveresultsthatitisgeneticallymodified.Ourstudyconfirmedthatcornis

geneticallymodified,whichgoesalongwithwhatisalreadyknown.Itisimportantforpeopleto

knowwhatisgeneticallymodifiedbecausesomedonotwanttoconsumetheseproducts.There

isagapbetweenpeoplethatknowaboutgeneticallymodifiedfoodandthosewhodonot

becausethereisalotofinformationthatcansometimesbehardtogetthrough(Mahgoub,2015).

Labelinggeneticallymodifiedfoodsisagoodwaytoletpeopleknow,butsinceonlyafewstates

intheUnitedStatesrequirelabeling,theinformationneedstobespreadotherways(Jalonick,

2015).

Agoodfutureexperimentcouldbeonethatanalyzesfoodsthatarenotcommonly

geneticallymodified.Theexperimentcouldlookatfoodsthatdonotsayiftheyaremodifiedor

iftheyarenot.Thisexperimentwasagoodstartingpointbecausewegottheresultswewere

supposedtoandthestudyworkedforus.So,wecouldfurthertheresearchwithotherfoodsand

crops.

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References

Bradshaw,J.E.(2016).GeneticallyModifiedCrops.InPlantBreeding:Past,Present,and

Future(Ch17).Retrievedfrom

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GreenFacts(n.d.).GeneticallyModifiedCrops.Retrievedfrom

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Havahart(n.d.).WhyOrganic:HarmfulEffectsofChemicalPesticides.Retrievedfrom

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Jalonick,M.C.(2015).HousePassesBilltoPreventMandatoryGMOFoodLabeling.PBS.

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http://www.upennsynapse.com/uploads/3/1/7/8/31787151/1081.pdf#page=30

Mahgoub,S.E.O.(2015).GeneticallyModifiedFoods:Basics,Applications,andcontroversy.

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