GeneticallyModifiedPlants

andAnimals

TheManipulationofGenes
y Gene asegmentofDNAina

chromosomespecifyingaparticular
proteinorpolypeptidechain,a
tRNAoranrRNA
y RecombinantDNA any
artificiallycreatedDNAmolecule
whichbringstogetherDNA
sequencesthatarenotusually
foundtogetherinnature.(Primrose
&Twyman,2006)

TheManipulationofGenes
(contd)
y GeneManipulation avarietyofsophisticated

techniquesforthecreationofrecombinantDNA,
whicharethenintroducedintolivingcells.
(Primrose&Twyman,2006)
y GeneticEngineering theisolation,
manipulation,recombination,andexpressionof
DNAoftenforthedevelopmentofgenetically
modifiedorganisms.

FindingtheRightGene
y Allgeneticengineeringstartsbyidentifying

andisolatingthecorrectclonecontaining
thegene
y Thiscanbedoneby:
y Makinggenelibrariesfromtotalgenomic
DNA
y Or,ifthegeneisidentified,cloningthe
DNAfragmentbyPCR(polymerasechain
reaction)

GeneLibraries
cDNALibraries
Theselibrarieswill
onlycontainDNA
fromtranscribed
genes.
GenomicDNA
Libraries These
librarieswillcontain
allDNAsequences

PolymeraseChainReaction(PCR)
Step1

PCR Step2

PCR Step3

PCR Step4

CuttingtheDNA
y Onceidentified,thenextstepisto

removethegeneyouareinterestedin
fromitshostorganism.
y TocuttheDNA,substancescalled
restrictionenzymesareused.
Restrictionenzymescutatspecific
locationsasdeterminedbytheDNA
sequence.

CuttingtheDNA (contd)
y Whenusingbacteria,thecutsegmentisthen

insertedintoasmall,circularpieceofbacterial
DNA,calledaplasmid.
y TheenzymeDNAligasesealsthebondbetween
thetransferredgeneandtheplasmidDNA
DNAligaserepairing
chromosomaldamage.
(ImagecourtesyoftheNational
InstitutesofHealth.)

GrowingtheGene
y Theplasmidisthenmixedwiththebacteriaand

spreadontoagrowthmediuminaPetridish.

GrowingtheGene(contd)
y Manyofthebacteriawillpickupthe

plasmid.
y Todeterminewhichbacteriapossessthe
newgene,specificmarkerssuchas
antibioticresistanceareinsertedalongwith
thegene.
y Thegrowthmediacontainsthetarget
antibiotic;thereforeorganismswhichgrow
onthemediummustcontainthenewgene

UsingtheNewGene
y Themainreasonforcreatinganewgeneistoproduce

theprotein.
y Someproteinsareusedtomakeplantsthatareresistant

toinsectsorinsecticides
y Someproteinsareusedtoenhancethecharacteristicsof

theproduct

FlavrSavrTomato

y Someproteinsareusedtomakepharmaceuticals

Insulin

GeneticallyModifiedAnimals
(TransgenicAnimals)
y TheFDACenterforVeterinaryMedicine(CVM)

regulatesgeneticallyalteredanimalproducts
y Currentlynotransgenicanimalshavebeen

approvedforhumanconsumption
y Transgenicanimalshavebeenapprovedforuseas

biopharmanimals(forproducingdrugsand
hormones)andtheyproducesuchproductsas
milkandwool

MakingTransgenicAnimals
y Makingageneticallymodifiedanimalcan

bedoneby:
y DNAmicroinjection Thenewgeneisinserteddirectly

intothefertilizedovum

MakingTransgenicAnimals(contd)
y Retrovirusmediatedgenetransfer RNAvirusesare

usedtotransferthegeneintothecell

MakingTransgenicAnimals(contd)
y EmbryonicStemCellMediatedGeneTransfer

thegeneisinsertedintoembryonicstemcells
soonafterfertilizationandthenimplantedinto
surrogatemothers

MouseEmbryo

EmbryonicStemCellMediatedGeneTransfer

Thismethodworks
verywellinmice
producingthe
knockoutmice
usedforlaboratory
research

BenefitsofTransgenicAnimals
y Productionofanimalswithspecifictraits

muchquickerthanwithtraditional
breedingmethods
y Resultsin
y Betterqualityandincreased
milkproduction
y Betterqualityandincreased
woolproduction
y Increasedgrowthrates

BenefitsofTransgenicAnimals(contd)
y Efficientproductionofpharmaceuticals,

nutritionalsupplements,andhormones
y Mostpharmaceuticalsareproducedfromthe

milkofgoats,cowsandsheep
y

Includedaresuchdrugsas
y
y

Insulin
Growthhormone

BenefitsofTransgenicAnimals(contd)
BiopharmAnimalsReduceProductionCosts
y

Expertsestimatethatproducingtherapeutic
proteinusingtraditionalmethodscost
approximately$300 $3,000pergram.
Incontrast,usingatransgenicgoattoproducethe
proteininmilkcostsapproximately$20 $105per
gram
Transgenicheneggsareevencheaper,costing
approximately$.10 $.25pergramofprotein

GeneticallyModifiedPlants
y Plantsaregeneticallymodifiedtobe:
y Herbicideresistant
y Pesticideresistant
y Insectresistant
y Droughttolerant
y Extremetemperaturetolerant
y Haveaddednutrients,suchasvitaminsandminerals

Roundupreadycrops
y Roundupisacommonherbicidemanufacturedby

Monsantothatisharmfultoweedsandplantsalike
y Forthisreason,Monsantodevelopedalineof
Roundupreadycropsthatareresistanttothe
herbicide
y Byinsertinggene5enolpyruvylshikimate3
phosphate(EPSP)fromthebacteriaAgrobacterium,
plantssuchascorn,soybeans,cotton,andalfalfacould
bemadeherbicideresistant

Insectresistance
y Corn,cotton,andseveralotherplantshavebeen

geneticallymodifiedtobeinsectresistant.
y Insectresistanceincropsisaccomplishedby
identifyingandisolatingagenefromthesoil
bacteriumBacillusthuringiensis thatproducesatoxin
calledCry thatistoxictoplantinsects.
y BycuttingandinsertingthegeneofinterestfromB.
thuringiensis intoplantDNA,anewgenetically
modifiedplantiscreatedthatisresistanttoinsects.

Benefitsofinsectresistance
y GMcottonplantsthatareinsectresistantare

protectedfromtobaccobudworm,bollworm,andpink
bollwormcaterpillars
y GMcornplantsthatareinsectresistantareprotected
fromEuropeancornborersandcornrootworms
y Theseinsectscauseseveredamagetotheplantsand
ultimatelyprovecostlytothefarmer.
y GMplantsarebeneficialinthattheyincreasecropyield

andsavefarmerstimeandmoneybyhavingtouseless
insecticides

GMplantswithaddednutrients
Rice
y Rice,astaplefoodinmanycountries,hasbeen

geneticallymodifiedtobeanimprovedsourceof
vitaminA
y ThisGMriceisabletobiosynthesizecarotene,
whichleadstoproductionofvitaminAinthehuman
body
y BiosynthesisofbetacaroteneinGMricewas
accomplishedbyinsertingphytoenesynthase(psy)
genefromdaffodilsandphytoenedesaturase(ctrI)
genefromthebacteriaErwiniauredovora intorice
DNA

Goldenrice
y Theadditionalbetacaroteneproducedbythe

endosperm(ricegrainthatiseatenbythehumans)
givesitacharacteristicyelloworgoldenhue
y BecauseofthisthevitaminenrichedGMriceisalso
knownasgoldenrice

White, non-GM rice

Golden, GM rice

GMplantswithaddednutrients
Strawberries
y Strawberries,whichareagoodsourceofvitaminC,havebeen

geneticallymodifiedtoprovide3timesasmuchvitaminC
y AgeneinthestrawberryplantcalledGalURgenecodesforan
enzymethatconvertsaproteinintheplanttovitaminC
y AsimilargeneisfoundinthethalecressArabidopsisthaliana.
y ResearcherscreatedaDNAplasmidusingtheA.thaliana gene
andthebacteriaAgrobacterium andinsertedintothestrawberry
planttooverexpressGalURgeneandproduce3timesasmuch
vitaminC

MoreGMcrops
y Currently,researchersaroundtheworldareworkingat

creatingandperfecting:
y Droughtresistantwheat,corn,andrice
y Salttoleranttomatoes
y Frostresistantstrawberries
y Heattolerantbeanssuchaskidney,red,black,andpinto

beans
y CarrotsthatproduceavaccineagainsthepatitisB

BenefitsofGMplants
y GMplantscould:
y Provideadditionalnutrients
y Resistinsects,herbicides,anddiseases
y Tolerateenvironmentalstressestoprovideanincreased
cropyield
y Provideenoughfoodforthegrowingpopulation
y Beasourceofvaccinesanddrugsforinfectiousdiseases

References
y
y
y
y
y
y
y
y

U.S.FoodandDrugAdministration.GuidanceforIndustryRegulationofGeneticallyEngineered
AnimalsContainingHeritablerDNA Constructs.http://www.fda.gov/cvm/Guidance/guide187.htm
NationalHumanGenomeResearchInstitute.http://www.genome.gov/12514551
Primose,S.B.andTwyman,R.M.PrinciplesofGeneManipulationandGenomics,7thed.2006.
BlackwellPublishing,Malden,MA.
Margawati,E.TransgenicAnimals:TheirBenefitsToHumanWelfare.Actionbioscience.org
publication.http://www.actionbioscience.org/biotech/margawati.html
Buy,M.TransgenicAnimals.CCACResourceSupplement,Spring/Summer1997.
http://www.ucalgary.ca/~browder/transgenic.html
Lewcock,A.Downonthebiopharm http://www.inpharmatechnologist.com/Industry
Drivers/Downonthebiopharm
http://www.actionbioscience.org/biotech/margawati.html
Lundmark,C.December2007.GeneticallyModifiedMaize.Bioscience57(11):996.Availableat:
http://web.ebscohost.com.proxy
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e5a7f1b86f83%40sessionmgr102&bdata=JmxvZ2lucGFnZT1Mb2dpbi5hc3Amc2l0ZT1laG9zdC1saXZl#d
b=aph&AN=28055306.
Widhalm,S.January2006.ProsandConsofTinkeringWithCropGenetics.World&I21(1):8.
Availableat:http://web.ebscohost.com.proxy
um.researchport.umd.edu/ehost/detail?vid=2&hid=116&sid=c8c74fe7159749fcad9a
0c6a7cbc2fad%40sessionmgr103&bdata=JmxvZ2lucGFnZT1Mb2dpbi5hc3Amc2l0ZT1laG9zdC1saXZl#d
b=ulh&AN=21419861

References(contd)
y
y
y
y

VanDeynze A.2004.RoundupReadyAlfalfa:AnEmergingTechnology.Availableat:
anrcatalog.ucdavis.edu/pdf/8153.pdf
Sawahel W.October14,2004.EgyptianscientistsproducedroughttolerantGMwheat.Availableat:
http://www.scidev.net/en/news/egyptianscientistsproducedroughttolerantgmwh.html
ScienceNews.November30,2007.BreedingHeatTolerantBeanstoWithstandWarmerWorld.
Availableat:http://www.sciencedaily.com/releases/2007/11/071126152058.htm
YeX,AlBabili S,Kloti A,ZhangG,LuccaP,BeyerP,Potrykus I.January14,2000.Engineeringthe
Provitamin A(Carotene)BiosyntheticPathwayinto(Carotenoid Free)RiceEndosperm.Science
287(5451):303.Availableat:http://www.sciencemag.org.proxy
um.researchport.umd.edu/cgi/content/full/sci;287/5451/303?maxtoshow=&HITS=10&hits=10&RESUL
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Magner L,GoldsteinN,FlanneryM.2004.Aregeneticallymodifiedfoodsandcropsdangerousto
humanhealthandtotheenvironment.ScienceinDisputeVol 3.Availableat:
http://findarticles.com/p/articles/mi_gx5204/is_2004/ai_n19124344/pg_11
Hancook R.March15,2006.ImprovingtheNutritionalValueofCropsbyGeneticModification:
ProblemsandOpportunitiesIllustratebyVitaminC.AsiaPacificBiotechNews10(5):237.Available
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um.researchport.umd.edu/ehost/pdf?vid=2&hid=108&sid=a106f370a23043afa315
2379943d7a8f%40sessionmgr109

PhotoReferences
y
y
y
y
y
y
y
y
y
y
y
y
y
y

DNAmicroinjection http://www.biologyreference.com/images/biol_01_img0084.jpg
GeneLibrary http://www.emunix.emich.edu/~rwinning/genetics/tech4.htm
Insulin http://www.diabetesmonitor.com/gifs/lantus.jpg
Tomato http://www.lhup.edu/smarvel/Seminar/FALL_2003/Malawskey/tomaten.jpg
Riceplant http://camereye.com/images/2005050201485112_rice.jpg
Petridish http://www.nasa.gov/centers/ames/images/content/173697main_ssbrp.jpg
DNAligase http://ocw.mit.edu/NR/rdonlyres/BiologicalEngineering/20450Spring
2006/6477D85A2C864B198E09DDDE66E9C4F4/0/chp_dnarepair.jpg
ViralGeneTransfer http://www.genecure.com/images/image004.gif
ESTransfer http://nobelprize.org/nobel_prizes/medicine/laureates/2007/advfig
1_web.gif
Mouseembryo http://www.albany.edu/genomics/graphics/mouse_embryo2.jpg
Cow http://www.ars.usda.gov/is/pr/2005/050404.D021_059i.jpg
Timemagazinecover http://openlearn.open.ac.uk/file.php/2808/S250_1_008i.jpg
Gene http://www.topnews.in/health/files/Genes.jpg
PCR http://library.thinkquest.org/24355/media/animations/polystep1.gif

PhotoReferencescontd
y CuttingDNA

y
y
y
y

http://dir.coolclips.com/Science/Chemistry/DNA/scientist_cutting_DNA_strand_wb031
896.html
GMcorn http://www.geocities.com/veronicaguseva/cornGM.jpg
Roundupanduntreatedcontrol anrcatalog.ucdavis.edu/pdf/8153.pdf
Goldenrice http://en.wikipedia.org/wiki/Image:GoldenRiceWhiteRice.jpg
Strawberries http://en.wikipedia.org/wiki/Image:Strawberries.JPG