Rgt Study Guide 4-1-2010

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RevisionDate:04.01.2010

RGT/CGT

STUDYGUIDEFORINFORMATIONORHELPFROMTHESCST,CONTACT:

AnitaHall

SCSTExecutiveDirector

101EastStateSt.,#214

Ithaca,NY14850

Ph:(607)2563313

Fax:(607)2731638

[email protected]

www.seedtechnology.net

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mailto:[email protected]://www.seedtechnology.net/http://www.seedtechnology.net/mailto:[email protected]

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TABLE OFCONTENTS

TableofContents_____________________________________________________________ 2Introduction_________________________________________________________________ 3MissionStatement__________________________________________________________________ 3

MembershipCategores______________________________________________________________ 3

RGT/CGTMembership_________________________________________________________ 4MembershipProcess________________________________________________________________ 4

ApplicationsforMembership_________________________________________________________ 5

MembershipQualifications___________________________________________________________ 5

ExaminationProcess________________________________________________________________ 6

PassingGrades_____________________________________________________________________ 6

ExaminationFormat________________________________________________________________ 7

MaintenanceofMembership ___________________________________________________ 8ContinuingEducation_______________________________________________________________ 8

PreparingfortheRGT/CGTExaminations__________________________________________ 8GeneralReferences:________________________________________________________________ 8

MolecularGeneticsReferences:_______________________________________________________ 9

ElectrophoresisReferences:__________________________________________________________ 9

ELISAReferences: _________________________________________________________________ 10

HerbicideBioassayReferences:______________________________________________________ 10

PCRbasedTechnologyReferences:___________________________________________________ 11

Glossary___________________________________________________________________ 11

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INTRODUCTION

MISSIONSTATEMENT

SCSTpromotesprofessionalismandensuresproficiencybyexaminingandcontinuingtoeducateseed

analysts.Thisprovidesaccurateandtimelyinformationtotheseedindustry.TheSCSTwillbuildupon

thesestrengthsbybroadeningthemembershipbasetoincludeemergingtechnologies.SCSTwill

continuetopromoteresearchanddeveloppublicationswhichenhanceseedtechnology.

TheSocietyofCommercialSeedTechnologistsisaseedtestingorganizationcomprisedofcommercial,

independentandgovernmentseedtechnologists.Formedin1922,theSCSTfunctionedasaliaison

betweentheAssociationofOfficialSeedAnalysts(AOSA)andtheAmericanSeedTrade(ASTA).TheSCST

hasdevelopedovertheyearsintoaprogressiveorganizationthattrainsandprovidesaccreditationof

technologists,researchesanddevelopsrulechanges,publishestrainingandeducationmaterials,and

serves

as

an

important

resource

to

the

seed

industry.

MEMBERSHIP CATEGORES

ThereareeightmembershipcategoriesintheSCST. Fiveofthemembershipcategories(RST,RGT,CGT,

CVT,CPT)requirequalifyingforandpassinganexamination. Researchmembershavetomeetcertain

qualificationsrelatedtoaccesstoresearchfacilitiesandresearchhistoryinordertobecomemembers.

Associatemembershipisopentoallindividualswithaninterestinseedtesting

Dividedintoeightcategories:

1.

Registered

Seed

Technologist

2.RegisteredGeneticTechnologist3.CertifiedGeneticTechnologist4.CertifiedViabilityTechnologist5.CertifiedPurityTechnologist6.ResearchMember7.AssociateMember8.HonoraryMember

REGISTERED GENETICTECHNOLOGIST (RGT)

AnRGThasqualifiedforandpassedthreeofthefourgenetictechnologyexamscurrentlyavailable:

herbicidebioassay,electrophoresis,immunoassay(ELISA),andPCR. TheRGTexamincludesarequired

writtenmoleculargenetics/biologyexamandareaspecificwrittenandpracticalexamsinthefour

genetictechnologyareas.Theseexaminationsaredesignedtoestablishtheapplicantscompetencyasa

welltrainedgenetictechnologistintheseedindustry.

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RGTsarerequiredtocompletecontinuingeducationinordertomaintainmembershipandarerequired

topayannualmembershipdues. TheymustsignaMembershipContractforPrivilegeofUseofthe

Societysname,logo,RGTseal,andthetitleRegisteredGeneticsTechnologist.

RGTshaveonevoteonallSocietybusinessandcanvoteontheamendmentstotheAOSARulesfor

Testing

Seeds.

RGTs

are

eligible

to

run

for

elected

office

and

can

chair

or

participate

on

committees.

CERTIFIEDGENETICTECHNOLOGIST(CGT)

ACGThasqualifiedforandpassedoneortwoofthefourgenetictechnologyexamscurrentlyavailable:

herbicidebioassay,electrophoresis,immunoassay(ELISA),andPCR. TheCGTexamincludesarequired

writtenmoleculargeneticsandareaspecificwrittenandpracticalexamsinthefourgenetictechnology

areas.Theseexaminationsaredesignedtoestablishtheapplicantscompetencyasawelltrained

genetictechnologistintheseedindustry.

CGTsarerequiredtocompletecontinuingeducationinordertomaintainmembershipandarerequired

topayannualdues.TheymustsignaMembershipContractforPrivilegeofUseoftheSocietysname,logo,andthetitleCertifiedGeneticTechnologist.

CGTshaveonevoteonallSocietybusinessandcanvoteontheamendmentstotheAOSARulesforTestingSeeds. CGTsareeligibletorunforelectedofficeandcanchairorparticipateoncommittees.

RGT/CGT MEMBERSHIP

MEMBERSHIP PROCESS

MembershipasaRegisteredGeneticTechnologist(RGT)orCertifiedGeneticTechnologist(CGT)shallbe

attainedinthefollowingorder:

1. CompleteandreturnthemembershipapplicationbyMarch1stannually. ApplicationsareavailablefromtheSCSTwebsite:http://www.seedtechnology.net/Membership.htm

2. Fulfillthequalificationsformembershipasprescribed(accumulationof100points)atleast14dayspriortowrittenexaminationday.ThewrittenexaminationwillbegivenduringSocietyof

CommercialSeedTechnologist(SCST)annualconference.

3. Beactivelyinvolvedingenetictestingforaminimumofoneyear.4. ObtainunanimousapprovaloftheRGTBoardofExaminers(BOE). Ifaunanimousvoteofsaid

Boardcannotbeobtained,theSCSTExecutiveBoardwillactasaBoardofReview.

5. Attainpassinggradesintheprescribedwrittenandpracticalexaminations.Theexaminationsconsistofdemonstratedwrittenandpracticalcompetencyineachofthefourareasofgenetic

testing: bioassay(herbicide),EnzymeLinkedImmunosorbentAssay(ELISA),electrophoresis

protocolsandpolymerasechainreaction(PCRbased)technologies.

a. AnindividualpassingthewrittenandpracticalportionsforanyofthefourareasoftheexaminationwillbeconferredthetitleofCertifiedGeneticTechnologistinthatarea(s)

ofcompetency.
http://www.seedtechnology.net/Membership.htmhttp://www.seedtechnology.net/Membership.htmhttp://www.seedtechnology.net/Membership.htm

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b. AnindividualpassingthewrittenandpracticalportionsofthreeofthefourareasoftheexaminationwillbeconferredthetitleRegisteredGeneticTechnologist.

6. Payanyduesorassessments.7. SignandreturnRGTorCGT MembershipContract

APPLICATIONS FORMEMBERSHIP

MembershipapplicationscanbedownloadedfromtheSCSTwebsiteorareavailablefromtheExecutive

Director. PleasechecktheSCSTwebsiteorcontacttheExecutiveDirectortoensurethatyouare

submittingacurrentversionofthemembershipapplication. Itisrecommendedthatyousubmitatrial

applicationtotheExecutiveDirectorbeforetheMarch1stdeadlinetoensurethatyouhavefulfilledthe

requirementsformembership. PleasecontacttheExecutiveDirectorifyouhaveanyquestionsorneed

helpcompletingtheapplicationformembership

MEMBERSHIP QUALIFICATIONS

Applicantsapplyingformembership,asaRGTorCGTshallmeetthefollowingqualifications:

POINTS REQUIRED TO QUALIFY FOREXAMS

InordertoqualifytotaketheRGT/CGTexamthecandidatemustaccumulateaminimumof100points

fromworkexperience,workshops/meetings,andcollegecourses.

1. Collegecourses:AcceptedaccreditedcoursesinBiologicalandMolecularSciences3pointsforeachearnedsemesterhour(2pointsforeachearnedquarterhour). Maximumof50pointsallowed.

Examples

of

accredited

courses

which

will

be

accepted:

PlantPhysiology PlantTaxonomy PlantGenetics PlantBreedingMolecularGenetics Biotechnology Biochemistry ChemosystematicsResearchMethods GeneExpression PlantGenome CellBiologyBiometrics Bacteriology Microbiology HorticultureBiology Immunology

2. ApprovedGeneticPurityWorkshops Maximumof20points.Note: Anadditional5pointswillbeallowedinthiscategoryforfullattendanceatanSCSTAnnualConference. (Priortotakingtheexamination)

Workshoppointswillbecreditedat2points/fulldayand1point/halfdaywhentheyaredirectlyrelatedtogeneticpuritytestingcomprisingaminimumof50%oftimetowardhandsontypeprogram. Workshopscomprisinglessthan50%oftimetowardshandsonexperiencewillbecreditedat1point/fullday.

8. WorkExperience. Candidatemustbeactivelyinvolvedingenetictestingforaminimumofoneyear. Trainingunderthesupervisionofaqualifiedsupervisor(RGT,CGT,orotherRGTBOE

approvedindividual). 1pointforeach40hourstraining. Unsupervisedgeneticpuritytesting

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experience:1pointforeach80hoursexperience.Combinationofthesewhichmeetthe

requirementofaminimumofoneyearofexperienceinhandsongeneticpuritytesting.

IfhandsongeneticpuritytestingexperiencewasobtainedearlierthantheimmediateoneyearpriortosubmittingapplicationforRGTorCGTexamination,applicantshallcompletethefollowingadditionalrequirement: Note: ProofoffivepointsofhandsoncontinuingeducationbetweenthetimeoforiginaltrainingandapplyingfortheRGTorCGTexamination.

EXAMINATION PROCESS

1. SubmitawrittenapplicationformtotheSCSTExecutiveDirectorincludingapplicationfeebyMarch1st.

2. ApplicantsareapprovedbytheRGTBOE,IfaunanimousvoteoftheRGTBOEcannotbeobtained,theExecutiveBoardwillactasaBoardofReview.

3. CandidatestakethewrittenexaminationattheSCSTannualmeetingandtheRGTBOEchairinforms

the

candidate

of

their

results

at

that

time.

Candidates

must

pass

the

written

examinationportion(s)beforeapplyingforthepracticalexaminationportion(s).Writtenexams

willnotbereturnedtotheexaminee.

4. ApplytotheSCSTExecutiveDirectorusingtheGeneticTechnologistPracticalExamApplicationform. Onthisformthecandidateselectswhichofthefourareastheyarerequestingsamples

forandtherespectivespecies/methods. Thecandidatemustalsodesignateaproctor.

5. TheapprovedproctorwillreceivethesamplesfromtheExecutiveDirectorandinformthecandidatethatsamplesareonsite. Theproctorwillusetheproctorformstorecordreceiptof

samples,datecandidatestartspractical,andverifyrequiredstepsorpointsofinspectioninthe

practicalexaminationprocess. SampleresultsmustbereturnedtoSCSTExecutiveDirector

within30daysfromthesamplesbeingtransferredtotheexaminee.Controlsamplesmaybe

giventoexamineepriortotestsamples

6. Oncetheexamination(s)arecompletedtheproctorwillmail/fax/emailtheresultsandproctorformstotheExecutiveDirector. TheExecutiveDirectorwillforwardcodedpracticalexamsto

therespectiveBOEmembersforgrading.

7. Practicalexamscoreswillbereturnedtothecandidatewith30daysofreceiptbytheExecutiveDirector. Practicalexamswillnotbereturnedtotheexaminee.

8. AnindividualpassingthewrittenandpracticalportionsforanyofthefourareasoftheexaminationwillbeconferredthetitleofCertifiedGeneticTechnologistinthatarea(s)of

competency. Anindividualpassingthewrittenandpracticalportionsofthreeofthefourareas

oftheexaminationwillbeconferredthetitleRegisteredGeneticTechnologist.

PASSINGGRADES

1. AllgradingoftheexaminationisbyRGTBOEmembers. Alltestsareidentifiedbynumberonly,notbyname.

2. PassingGrades:Anapplicantshallfulfillthefollowingqualifications

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a. Achieveagradeof70%orbetteroneachpartofthewrittenandpracticalexamination.b. Achieveanaveragegradeof80%orbetterforthewrittenandpracticalexamsineach

areainwhichcertificationissought.

Example:inordertobecomecertifiedinherbicidebioassaytestingthemoleculargenetics

written

exam

score

will

be

averaged

with

the

herbicide

bioassay

written

and

practical

exam

scores.Eachindividualscoremustbe70%orhigherandthetotalscoremustbe80%or

higher

EXAMINATION FORMAT

WRITTEN EXAMINATIONS

1. WrittenexaminationsareconductedbytheRGTBOEattheSCSTannualmeeting.2. Thewrittenportionoftheexaminationwillconsistof:

a. Ageneralmoleculargenetics/biologyexam takenbyallcandidatesb. FourareaspecificexamsemphasizingBioassay,ELISA,ElectrophoresisandPCRbasedtechniques.Onehourandfifteenminutesaregivenforeachofthefiveexams. Breaks

aregivenbetweeneachexam.

3. Questionswillbeacombinationofmultiplechoice,true/false,shortanswer,andessaytypes.CandidateswillcompletetheexamsandbeinformedoftheirscoresattheSCSTannualmeeting.

PRACTICAL EXAMINATIONS

1. Practicalexamscanonlybetakenafterthewrittenexamshavebeenpassed.2.

The

practical

portion

of

the

examination

will

consist

of

four

parts

emphasizing

Bioassay,

ELISA,

ElectrophoresisandPCRbasedtechniques.Examinationsmaybetakenwithinthecandidates

laboratory,oratanapprovedlocation.

3. OncecompletedresultsshallbeimmediatelyturnedintotheExecutiveDirector.ProctoredexaminationsshallbegradedbytherespectiveRGTBOEmembers,returnedtotheExecutive

Directorandthecandidatewillbenotifiedoftheirtestscore.

4. Practicalexaminationsmustbecompletedwithinoneyearfromthetimethecandidatepassesthewrittenexamination. TheExecutiveDirectorwillsendquarterlyreminderstocandidates.

5. Ifacandidatefailsthefirstpracticalexaminationtheymustcompleteeighthoursofcontinuingeducation. Acceptablecontinuingeducationincludes:

a. IndividualizedstudywithaBOEapprovedtutor. AnagendamustbesubmittedtotheExecutiveDirectorforapprovalpriortothetrainingsession.b. Attendanceatagenetictechnologyworkshop.c. OtherindependentstudyapprovedbytheRGTBOE.

6. Candidatesmustwaitaminimumoftwomonthsbeforeretakingtheexam. Onceapprovedtoretakethepracticalexamitmustbecompletedandreturnedwithinfourmonths. Ifthe

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candidatefailstheexamagaintheywillberequiredtoretakeboththewrittenandpractical

exam.

PRACTICAL EXAMINATION PROCTORS

1. Examproctorsareusedtomonitorcandidatestakingoneormoreofthepracticalexams.2. AnacceptableproctormaybeanRST,RGT,orCGT. TheRGTBOEwillapproveotherproctorson

acasebycasebasiswhenneeded.

3. Theroleofproctoristomakesurethecandidateisdoingtheworkbutwillnotgradeorassistthecandidateintakingtheexam. Proctorsreceivetheseedsamplesandtestforms,deliver

themtothecandidate,andmonitortestingofcriticalstepsforeachsection. Whentestingis

complete,theproctorsendsthetestmaterialstotheexecutivedirector.

MAINTENANCE OF MEMBERSHIP

CONTINUING

EDUCATION

AllRGTandCGTmembersarerequiredtocomplete5pointsofcontinuingeducationeverythreeyears:1. Attendaminimumofthree(3)fulldaysattheAnnualMeetingoftheSociety,whichshall

includeattendanceattheSCSTbusinessmeeting,RegisteredorCertifiedMemberspresentatthemeetingbutnotinattendanceduringRollCallareresponsibleforhavingtheirnamerecordedbytheExecutiveDirector.

2. Attainfive(5)pointsforattendanceatworkshopsorseedschoolsdirectlyrelatedtoseedtestingthatcompriseahandsontypeprogramandhavebeenapprovedpriortoattendancebytheExecutiveDirector.

3. AttendindividualizedseedtechnologytrainingthatreceivespriorapprovalbytheExecutiveDirector. Pointsarecreditedonthebasisofone(1)pointforeverythree(3)hours,maximum

two

points

per

day.

A

certificate

of

attendance

must

be

submitted

to

the

Executive

Director

to

receiveproperpointcredits. Collegecreditsfromapprovedseedrelatedcourseswouldbeacceptableforuptohalf(1/2)ofrequiredpointsbasedonthree(3)pointsforeachsemesterhourortwo(2)pointsforeachquarterhour.

PREPARINGFORTHE RGT/CGT EXAMINATIONS

InadditiontothereferenceslistedinthissectionthereareanumberofresourcesavailableontheSCSTwebsiteathttp://www.seedtechnology.net/genetic_resources.htm. ThiswebpageincludespresentationsfrompastGeneticTechnologyWorkshops,TheLibraryofCropTechnologyLesson

Modules

from

the

University

of

Nebraska

Lincoln,

and

other

helpful

websites.

Workshops

and

training

opportunitiesarealsolistedonthewebsite.

ItishighlyrecommendedthatcandidatesreviewthepresentationsfrompastGeneticTechnology

Workshops;theseworkshopsarespecificallyfocusedonpreparationfortheRGT/CGTexams.

GENERAL REFERENCES:

1. SeedTechnologistTrainingManual.SCST2001.Chapter14GeneticPurityTestingupdate2005
http://www.seedtechnology.net/genetic_resources.htmhttp://www.seedtechnology.net/genetic_resources.htm

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2. CultivarPurityTestingHandbook. AOSA,updated20083. HandbookofVarietyTesting. GrowthChamberGreenhouseTestingProcedures: Variety

Identification. ISTA19934. HandbookofVarietyTesting. LaboratoryTestsforVarietyDeterminationwithFungal

Pathogens. ISTA1993.5. HandbookofVarietyTesting. RapidChemicalIdentificationTechniques. ISTA1993.6. Singer,M.,P.Berg,1991. GenesandGenomes.UniversityScienceBooks.7. Smith,J.S.C. 1992. PlantbreedersrightsintheUSA;changingapproachesandappropriate

technologiesinsupportofgermplasmenhancement. PlantVar.Seeds.5:183199.8. Wrigley,C.W. 1995. IdentificationofFoodGrainVarieties. Amer.Assoc.CerealChem.,St.Paul,

MN. 283pp9. McGrawHillDictionaryofScientificandTechnicalTerms,FifthEdition.10.OxfordDictionaryofBiochemistryandMolecularBiology. OxfordUniversityPress11.GlossaryofBiotechnologyTermshttp://biotechterms.org12. ISTAStatisticalToolBoxhttp://www.seedtest.org/en/stats_tool_box_content11143.html

MOLECULAR GENETICSREFERENCES:

1. Weaver,RobertF.2002. MolecularBiology,2nd edition.TransmissionGenetics,glossary.NewYork,McGrawHillCo.,Inc.

2. Lewin,Benjamin..GenesIX3. Klug,WilliamS.andCummings,MichaelR.ConceptsofGenetics

ELECTROPHORESIS REFERENCES:

1. Andrews,A.T.1995.Electrophoresis Theory,Techniques,andBiochemicalandClinicalApplications,2

ndedition,Introduction. Oxford,NY:ClarendonPress.

2. BourgonGreneche,M.G.Giraud,R.Pouget.1994.Technicalreferencemanualfortheisoenzymaticanalysisofmaize.BIOGEVESLaboratoire.

3. Cardy,B.J.,C.W.Stuber,J.F.Wendel,andM.M.Goodman. 1983. Techniquesforstarchgelelectrophoresisofenzymesfrommaize(ZeamaysL.). N.C.StateUniv.Inst.Stat.MimeoSer.No.1317. 35pp.

4. Cooke,R.J. 1995. Gelelectrophoresisfortheidentificationofplantvarieties. J.Chromat.698:281299.

5. DombrinkKurtzman,M.A.andJ.A.Bietz.1993.ZeinCompositioninHardandSoftEndospermofMaize.CerealChem.70(1):105108

6. Goodman,M.M.andC.W.Stuber,1980.Geneticidentificationoflinesandcrossesusingisoenzymeelectrophoresis. Proceedings35thAnnualCornandSorghumIndustryResearch

Conference.7. McDonald,M.B. 1995. Geneticpurity: FromproteinelectrophoresistoRAPDs. Proc.Ann.Corn

andSorghumConf.50:2562768. Motto,M.andF.Salamini.1979.EvaluationofGeneticPurityinHybridCorn(ZeamaysL.)Seed

ProductionThroughZeinIsoelectrophoreticPatterns.MaydicaXXIV(1979):223233.9. HandbookofVarietyTesting. ElectrophoresisTesting. ISTA1992.10.Smith,J.S.C.,andJ.C.Register,III. 1998. Geneticpurityandtestingtechnologiesforseed

quality: acompanyperspective. SeedSci.Res.8:285293.
http://biotechterms.org/http://www.seedtest.org/en/stats_tool_box_content---1--1143.htmlhttp://www.seedtest.org/en/stats_tool_box_content---1--1143.htmlhttp://www.seedtest.org/en/stats_tool_box_content---1--1143.htmlhttp://www.seedtest.org/en/stats_tool_box_content---1--1143.htmlhttp://www.seedtest.org/en/stats_tool_box_content---1--1143.htmlhttp://www.seedtest.org/en/stats_tool_box_content---1--1143.htmlhttp://biotechterms.org/

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11.Stuber,C.W.andM.M.Goodman.1983.Allozymegenetypesforpopularandhistoricallyimportantinbredlinesofcorn,ZeamaysL.USDAARSARRS17/August1983.

12.Stuber,C.W.,J.F.Wendel,M.M.Goodman,J.S.C.Smith.1988.Techniquesandscoringproceduresforstarchgelelectrophoresisofenzymesfrommaize(ZeamaysL.)N.C.StateUniv.InstStat.MimeoSer.No.286.

13. Westermeier,Reiner.1997.ElectrophoresisinPractice,2ndedition.VCH,AWileyCo.14.Wilson,C.M.,et.al.1988.Linkagesamongzeingenesdeterminedbyisoelectricfocusing.

NorthernRegionalResearchCenter,USDAARSNationalCenterforAgricultureUtilizationResearch,1815N.UniversitySt.,PeoriaIL. TheorApplGenet(1989)77:217226.

15.Wilson,C.M.1984.IsoelectricFocusingofZeininAgarose.USDepartmentofAgriculture,AgricultureResearchService,1102S.Goodwin,Urbana,IL.CerealChem.61(2):198200.

Websitereferences:http://www.innvista.com/health/nutrition/amino/pclass.htm

ELISA REFERENCES:

1. Albertsetal.(1989)MolecularBiologyoftheCell2ndEd.GarlandPublishing,NewYork1218P2. Clark,M.F.,Lister,R.M.,andBarJoseph,M.(1988).ELISATechniques,MethodsforPlant

MolecularBiology. Pg.507529.3. Crowther,J.R.(1995). ELISA:Theoryandpractice.HumanaPress,Totowa,NJ223p1. Grothaus,David.(2006)ImmunoassayasanAnalyticalToolinAgriculturalBiotechnology.

JournalofAOACInternational,Vol.89,No.4 AvailablefordownloadfromtheSCSTLibraryWebpage:http://www.seedtechnology.net/Seed%20Library/rgt.html

2. Stave,James(2002).ProteinImmunoassayMethodsforDetectionofBiotechCrops:Applications,Limitations,andPracticalConsiderations. .JournalofAOACInternational,Vol.85,No.3AvailablefordownloadfromtheSCSTLibraryWebpage: http://www.seedtechnology.net/Seed%20Library/rgt.html

4. Sutula,C.L.(1996).QualityControlandCostEffectivenessofIndexingProcedures. AdvancesinBotanicalResearch,23,279292.

5. Voller,A.,Bidwell,D.E.,andBartlett,A.(1979). Thedetectionofvirusesbyenzymelinkedimmunosorbantassay(ELISA).J.Gen.Virol.33,165167. AvailablefordownloadfromtheSCSTLibraryWebpage: http://www.seedtechnology.net/Seed%20Library/rgt.html

HERBICIDE BIOASSAY REFERENCES:

1. Goggi,A.S.andM.G.Stahr.1997ROUNDUPpreemergencetreatmenttodeterminethepresenceoftheroundupreadygeneinsoybeanseed:alaboratorytest.SeedTechnology.

19(1):99102. AvailablefordownloadfromtheSCSTLibraryWebpage: http://www.seedtechnology.net/Seed%20Library/rgt.html

2. Gutormson,T.J.1999.BioassayprocedurefordeterminingthepresenceoftheRoundupReadygeneincorn.P1819.In:Testingmethodologiesforhybridparentageandtraitdetermination.Zaworkski,F.1999.Corn,Sorghum,andSoybeanTechnology1999.AspecialpublicationofSeedTradeNews.

3. Sebastain,S.A.andR.S.Chaleff.1987.Soybeanmutantswithincreasedtoleranceforsulfonylureaherbicides.CropSci.27:948952.
http://www.innvista.com/health/nutrition/amino/pclass.htmhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.seedtechnology.net/Seed%20Library/rgt.htmlhttp://www.innvista.com/health/nutrition/amino/pclass.htm

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AGROBACTERIUM TUMEFACIENS

AnaturallyoccurringbacteriumthatiscapableofinsertingitsDNA(geneticinformation)intoplants,resultinginatypeofinjurytotheplantknownascrowngall.In1980,MarcvanMontagushowedthatAgrobacteriumtumefacienscouldaltertheDNAofitshostplant(s)byinsertingitsown("foreign")DNAintothegenomeofthehostplants(therebyopeningthewayforscientiststoinsertvirtuallyanyforeign

genes

into

plants

via

use

of

Agrobacterium

tumefaciens).

Among

others,

Monsanto

Company

has

developedawaytostopAgrobacteriumtumefaciensfromcausingcrowngall,whilemaintainingitsabilitytoinsertDNAintoplantcells,andnowusesAgrobacteriumtumefaciensasavehicletoinsertdesiredgenesintoplants(e.g.,thegenecausingoverproductionofCP4EPSPsynthase,thusconferringresistancetoglyphosatecontainingherbicide).

ALLELE

FromtheGreekallelon="mutuallyeachother",thetermreferstooneofseveralalternateformsofageneoccupyingagivenlocusonthechromosome,whichcontrolsexpression(ofproduct)indifferentways.

ANTIBODY

Alsocalledimmunoglobulin,Ig.Alargedefenseproteinthatconsistsoftwoclassesofpolypeptidechains,light(L)chainsandheavy(H)chains.AsingleantibodymoleculeconsistsoftwoidenticalcopiesoftheLchainandtwooftheHchain.Theyaresynthesized(i.e.,made)bytheimmunesystem(Blymphocytes)oftheorganism.TheantibodyiscomposedoffourproteinslinkedtogethertoformaYshapedbundleofproteins(lookssomewhatlikeaslingshotortwohockeystickstapedtogetheratthehandles).Theaminoacidsequencethatmakesupthestem(heavychains)oftheY(i.e.,thehandlesofthetapedtogetherhockeysticks)issimilarforallantibodies.ThestemisknownastheFcregionoftheantibodyanditdoesnotbindtoantigen,butdoeshaveotherregulatoryfunctions.

ThetwoarmsoftheYareeachmadeupoftwosidebysideproteinscalledlightchainsandheavychains(i.e.,proteinsarechainsofaminoacids),withidenticalantigenbinding(ab)sitesonthetipsofeach"arm."Theantibodyisthusbivalentinthatithastwobindingsitesforantigen.Takentogether,thetwoarmsoftheYareknownastheFabportionsoftheantibodymolecule.TheFabportionscanbecleavedfromtheantibodymoleculewithpapain(anenzymethatisalsousedasameattenderizer)ortheFabportionscanbeproducedviageneticallyengineeredEscherichiacolibacteria.

Whenaforeignmolecule(e.g.,abacterium,virus,etc.)entersthebody,Blymphocytesarestimulatedintobecomingrapidlydividingblastcells,whichmatureintoantibodyproducingplasmacells.Theplasmacellsaretriggeredbytheforeignmolecule'sepitope(s)[i.e.,grouporgroupsofspecificatoms(alsoknownasahapten),thatarerecognizedtobeforeignbythebody'simmunesystem]intoproducingantibodymoleculespossessingantigenbinding(ab)sites(alsocalledcombiningsitesordeterminants).

Thesefitintotheforeignmolecule'sepitope.Thus,viathetipsofitsarms,theantibodymoleculebindsspecifically totheforeignentity(antigen)thathasenteredthebody.Bythisprocessitinactivatesthatforeignmoleculeormarksitforeventualdestructionbyotherimmunesystemcells.

Systemmarkingoftheforeignmolecule(e.g.,pathogenortoxin)fordestructionisaccomplishedbythefactthatthestemoftheY(i.e.,theFc)fragmenthangsfreefromthecombinedantibodyantigenclump,therebyprovidingareceptorforphagocytes,whichroamthroughoutthebodyingestingand

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subsequentlydestroyingsuch"marked"foreignmolecules.Thissystemiscalledantibodydependentcellularcytotoxicity.

Researchpublishedduring2001indicatesthatantibodiesmayalsokillsomepathogensthemselvesbycatalyzingtheformationofhydrogenperoxidefromoxygenfreeradicals(singletoxygen)andwater.Hydrogenperoxideishighlyreactive,andcouldpotentiallykillpathogenswhengeneratedbyan(attached)antibody.

Therearefiveclassesofimmunoglobulin:IgG,IgM,IgD,IgA,andIgE.

ANTIGEN

Alsocalledanimmunogen.Anylargemoleculeorsmallorganismwhoseentryintothebodyprovokessynthesisofanantibodyorimmunoglobin(i.e.,animmunesystemresponse).

ANTISENSE(DNASEQUENCE)

AstrandofDNAthatproducesamessengerRNA(mRNA)moleculewhich(whenreversedendforend)

has

the

same

sequence

as

(i.e.,

is

complementary

to)

the

unwanted

("bad")messenger

RNA.

The

SENSE

(i.e.,forward)andANTISENSE(i.e.,backward)mRNAstrandshybridize(i.e.,tightlybondtoeachother),whichpreventsthebondedpairfromleavingthecell'snucleus,sothatbodedpairisrapidlydegraded(destroyed)bynucleaseswithinthecellnucleus.

Ingenetictargetingusingantisensemolecules(toblock"bad"genes),antisensemoleculesareusedtobindtoa"bad"gene's(e.g.,anoncogene)messengerRNA(mRNA),thuscancellingthe(cancercausing)messageofthegeneandpreventingcellsfromfollowingits(tumorgrowth)instructions.AnotherexamplewouldbetheuseofantisenseDNAtoblockthegenethatcodesforproductionofpolygalacturonase(anenzymethatcausesripefruittosoften).

Physically,"antisense"isaccomplishedbyremovingagivengenefromanorganism'sgenome,reversing

it(endforend),andreinsertingitbackintotheorganism'sgenome.

ARABIDOPSISTHALIANA

Asmallweedplantpossessing70,000kilobasepairsinitsgenome,withverylittlerepetitiveDNA.Thismakesitanidealmodelforstudyingplantgenetics.AtleasttwogeneticmapshavebeencreatedforArabidopsisthaliana(oneusingyeastartificialchromosomes).Becauseofthisalargebaseofknowledgeaboutithasbeenaccumulatedbythescientificcommunity.

Arabidopsisthalianawasfirstgeneticallyengineeredin1986.In1994,researcherssucceededintransferringgenesforpolyhydroxylbutylate("biodegradableplastic")productionintoArabidopsisthaliana.Becauseproductionofpolyhydroxylbutylate(PHB)requiressimultaneousexpressionofthree

genes(i.e.,thePHBproductionprocessis"polygenic") yetresearchershaveonlybeenabletoinsertamaximumoftwogenes theyhavetoinserttwogenesintooneplantandonegeneintoasecondplant,thenfinallygetthe(total)threegenesinto(offspring)plantsviatraditionalbreeding.

ASSAY

Atest(specifictechnique)thatmeasuresaresponsetoatestsubstanceortheefficacy(effectiveness)ofthetestsubstance.

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BACILLUSTHURINGIENSIS (B.T.)

DiscoveredbybacteriologistIshiwataShigetaneonadiseasedsilkwormin1901.LaterdiscoveredonadeadMediterraneanflourmoth,andfirstnamedBacillusthuringiensis,byErnstBerlinerin1915.

Today,Bacillusthuringiensisreferstoagroupofrod shapedsoilbacteriafoundallovertheearth,that

produce

"cry"

proteins

which

are

indigestible

by

yet

still

"bind"

to

specific

insects'

gut

(i.e.,

stomach)

liningreceptors,sothose"cry"proteinsaretoxictocertainclassesofinsects(cornborers,cornrootworms,mosquitoes,blackflies,sometypesofbeetles,etc.),butwhichareharmlesstoallmammals.Atleast20,000strainsofBacillusthuringiensisareknown.

Genesthatcodefortheproductionofthese"cry"proteinsthataretoxictoinsectshavebeeninsertedbyscientistssince1989intovectors(i.e.,viruses,otherbacteria,andothermicroorganisms)inordertoconferinsectresistancetocertainagriculturalplants(e.g.,viaexpressionofthoseB.t.proteinsbyoneormoretissuesofthetransgenicplant)..Forexample,theB.t.strainknownasB.t.kurstaki,whichisfatal

wheningestedbytheEuropeancornborerwasfirst(genetically)insertedintoacornplant(viavector)in1991.B.t.kurstakikillsborersviaperforationofthatinsect'sgutbyproteinsthatarecodedforbytheB.t.kurstakigene.ThevectorsaslistedaboveareentitiesthatcantakeupandcarrytheDNAintoplantorothercells.VectorsareDNAcarryingvehicles.

BASEPAIR(BP)

Twonucleotidesthatareindifferentnucleicacidchainsandwhosebasespair(interact)byhydrogenbonding.InDNA,thenucleotidebasesareadenine(whichpairswiththymine)andguanine(whichpairswithcytosine). InRNA,thenucleotidebasesareadenine(whichpairswithuracil)andguanine(whichpairswithcytosine).

BIOASSAY

Determinationoftherelativestrengthorbioactivityofasubstance(e.g.,adrug).Abiologicalsystem

(such

as

living

cells,

organs,

tissues,

or

whole

animals)

is

exposed

to

the

substance

in

question

and

the

effectonthelivingtestsystemismeasured.

BIOCHEMISTRY

Thestudyofchemicalprocessesthatcompriselivingthings(systems).Thechemistryoflifeandlivingmatter.Despitethedramaticdifferencesintheappearancesoflivingthings,thebasicchemistryofallorganismsisstrikinglysimilar.Eventinyonecelledcreaturescarryoutessentiallythesamechemicalreactionsthateachcellofacomplexorganism(suchasman)carriesout.

BIOTECHNOLOGY

Themeansorwayofmanipulatinglifeforms(organisms)toprovidedesirableproductsforman'suse.

For

example,

beekeeping

and

cattle

breeding

could

be

considered

to

be

biotechnology

related

endeavors.Thewordbiotechnologywascoinedin1919byKarlEreky,toapplytotheinteractionofbiologywithhumantechnology.

However,usageofthewordbiotechnologyintheUnitedStateshascometomeanallpartsofanindustrythatknowinglycreate,develop,andmarketavarietyofproductsthroughthewillfulmanipulation,onamolecularlevel,oflifeformsorutilizationofknowledgepertainingtolivingsystems.

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AcommonmisconceptionisthatbiotechnologyrefersonlytorecombinantDNA(rDNA)work.However,recombinantDNAisonlyoneofthemanytechniquesusedtoderiveproductsfromorganisms,plants,andpartsofbothforthebiotechnologyindustry.Alistofareascoveredbythetermbiotechnologywouldmoreproperlyinclude:recombinantDNA,planttissueculture,rDNAorgenesplicing,enzymesystems,plantbreeding,meristemculture,mammaliancellculture,immunology,molecularbiology,fermentation,andothers.

BLANK (inanELISAplate)Measurestheopticaldensityassociatedwiththeregentsusedinthetestandplate. Awellwithnosamplethatisusedasabaselinefortheplatereader,removesanybackgroundcolor.

Btk

TransgenicproteinBacilluskurstakii,resistanttothecornborer.

CDNA

AlsoknownascopyDNA.AhelicalformofDNA.ItoccurswhenDNAfibersaremaintainedin66percent

relativehumidityinthepresenceoflithiumions.IthasfewerbasepairsperturnthanBDNA.

CARBOHYDRATES

(saccharides)Alargeclassofcarbonhydrogenoxygencompounds.Monosaccharidesarecalledsimplesugars,ofwhichthemostabundantisDglucose.Itisboththemajorfuelformostorganismsandconstitutesthebasicbuildingblockofthemostabundantpolysaccharides,suchasstarchandcellulose.Whilestarchisafuelsource,celluloseistheprimarystructuralmaterialofplants.Carbohydratesareproducedbyphotosynthesisinplants.Most,butnotall,carbohydratesarerepresentedchemicallybytheformulaCx(H20)n,wherenisthreeorhigher.Onthebasisoftheirchemicalstructures,carbohydratesareclassifiedaspolyhydroxyaldehydes,polyhydroxyketones,andtheirderivatives.

CATALYST

Anysubstance(entity),eitherofproteinorofnonproteinaceousnature,thatincreasestherateofachemicalreaction,withoutbeingconsumeditselfinthereaction.Inthebiosciences,theterm"enzyme"isusedforaproteinaceouscatalyst.Enzymescatalyzebiologicalreactions.

CAULIFLOWERMOSAICVIRUS35SPROMOTER (CaMV35S)Apromoter(sequenceofDNA)thatisoftenutilizedingeneticengineeringtocontrolexpressionof(inserted)gene;i.e.,synthesisofdesiredproteininaplant.

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stationaryphase.Chromatographyconstitutesoneof,ifnotthemostfundamentalseparationtechniquesusedinthebiochemistry/biotechnologyarenatodate.

CHROMOSOMES

Discreteunitsofthegenomecarryingmanygenes,consistingof(histone)proteinsandaverylong

molecule

of

DNA.

Found

in

the

nucleus

of

every

plant

and

animal

cell.

CLONE(ANORGANISM)

Agroupofindividualorganisms(orcells)producedfromoneindividualcellthroughasexualprocessesthatdonotinvolvetheinterchangeorcombinationofgeneticmaterial.Asaresult,membersofaclonehaveidenticalgeneticcompositions.Forexample,protozoaandbacteriafrequentlyreproduceasexually(i.e.,withoutsex)byaprocesscalledbinaryfission.Inbinaryfissionasinglecelledorganismundergoescelldivision.Theresultistwocellswithidenticalgeneticcomposition.Whenthesetwoidenticalcellsundergodivision,theresultisfourcellswithidenticalgeneticcomposition.Theseidenticaloffspringareallmembersofaclone.Theword"clone"maybeusedeitherasanounoraverb.

CODINGSEQUENCE

TheregionwithinaDNAmolecule(i.e.,betweenthestartandstopcodons)thatencodestheaminoacidsequenceofaprotein,orforaspecificmicroRNA.

COMPLEMENTARY DNA(cDNA)AsinglestrandedDNAthatiscomplementarytoastrandofmRNA.TheDNAissynthesizedinvitrobyanenzymeknownasreversetranscriptase.Then,asecondDNAstrandissynthesizedviatheenzymeknownasDNApolymerase.

ComplementaryDNAisoftenutilizedinhybridizationstudiesandinmicroarrays(e.g.,todetect/identifygenes)becausecDNAsusuallydon'tcontainregulatorysequencesofDNA;sincethecDNAwascopiedfrommRNA.BecausecDNAisaDNAcopyofmRNA(messengerRNA),itisanexceptiontothe(old)CentralDogma.

CONJUGATE

Amoleculecreatedbyfusingtogether(e.g.,viarecombinationorchemically)twounlike(different)molecules.Thepurposeofthisistocreateamoleculeinwhichoneoftheoriginalmoleculeshasonefunction,forexample,atoxic,cellkillingfunction,whiletheotheroriginalmoleculehasanotherfunction,suchastargetingthetoxintoaspecificsiteinthebody,whichmightbecancerouscells.

CONTAMINANT

Bydefinition,anyunwantedorundesiredorganism,compound,ormoleculepresentinacontrolled

environment.

Unwanted

presence

of

an

entity

in

an

otherwise

clean

or

pure

environment.

CP4EPSPS

Theenzyme5enolpyruvylshikimate3phosphatesynthase,whichisnaturallyproducedbyanAgrobacteriumspecies(strainCP4)ofsoilbacteria.CP4EPSPSisessentialforthefunctioningofthatbacterium'smetabolismbiochemicalpathway.CP4EPSPShappenstobeunaffectedbyglyphosatecontainingherbicides,sointroductionoftheCP4EPSPSgeneintocropplants(e.g.,soybeans)makesthoseplantsessentiallyimpervioustoglyphosatecontainingherbicides.

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DOMINANTALLELE

DiscoveredbyGregorMendelinthe1860s,itisagenethatproducesthesamephenotypewhenitisheterozygousasitdoeswhenitishomozygous(i.e.,trait,orprotein,isexpressedevenifonlyonecopyofthegeneispresentinthegenome).

ELECTROPHORESIS Atechniqueforseparatingmoleculesbasedonthedifferentialmovementofchargedparticlesthroughamatrixwhensubjectedtoanelectricfield.Thetermisusuallyappliedtolargeionsofcolloidalparticlesdispersedinwater.Themostimportantuseofelectrophoresis(currently)isintheanalysisofproteins,andthenatechniqueknownasgelelectrophoresisisused.Sincetheproportionofproteinsvarieswidelyindifferentdiseases,electrophoresiscanbeusedfordiagnosticpurposes.

Electrophoresis,throughagaroseorothergelmatrices,isacommonwaytoseparate,identify,andpurifyplasmidDNA,DNAfragmentsresultingfromdigestion(ofDNA)withrestrictionendonucleases,andRNA.Electrophoresisisalsousedtostudybacteriaandviruses,nucleicacids,andsometypesofmolecules,includingaminoacids.

ELISA(testforproteins).Anenzymelinkedimmunosorbentassay whichcanreadilymeasurelessthanananogram(109g)ofaprotein.Thisassayismoresensitivethansimpleimmunoassay(tests)becauseoneofthetwoantibodiesusedtobindandquantitate(measure)theprotein%antigen,basedontwoconcurrentepitopeswithintheprotein,isattachedtoanenzyme.Theenzymecanrapidlyconvertanaddedcolorlesssubstrateintoacoloredproduct,oranonfluorescentsubstrateintoanintenselyfluorescentproduct(thusenablingfinerquantitation).

ENZYME

Anorganic,proteinbasedcatalystthatisnotitselfusedupinthereaction.Itisnaturallyproducedbylivingcellstocatalyzebiochemicalreactions.Eachenzymeishighlyspecificwithregardtothetypeofchemicalreactionthatitcatalyzes,andtothesubstances(calledsubstrates)uponwhichitacts.Thisspecificcatalyticactivityanditscontrolbyotherbiochemicalconstituentsareofprimaryimportanceinthephysiologicalfunctionsofallorganisms.Althoughallenzymesareproteins,theymay,andusuallydo,containadditionalnonproteincomponentscalledcoenzymesthatareessentialforcatalyticactivity.

EPITOPE

Alsocalledantigenicdeterminant.Thespecificgroupofatoms(onanantigenmolecule)thatisrecognizedby(thatantigen's)antibodies.

ERRORRATE

Thepercentageoftimesthatthetestwillproduceafalsereading.

EVENT

Referstoeachinstanceofageneticallyengineeredorganism.Forexample,thesamegeneinsertedbymanintoagivenplantgenomeattwodifferentlocations(i.e.,loci)alongthatplant'sDNAwouldbeconsideredtwodifferent"events."Alternatively,twodifferentgenesinsertedintothesamelocusoftwosamespeciesplantswouldalsobeconsideredtwodifferent"events."

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Generallyspeaking,theworld'sregulatoryagenciesconfernewbiotechderivedproductapprovalsintermsofevents.

EXPRESS

Totranslatethecell'sgeneticinformationstoredintheDNA(gene)intoaspecificprotein

(synthesized

by

the

cell's

ribosome

system).

F1 HYBRIDS

Thefirstgenerationoffspringofcrossbreeding;alsoknownasfirstfilialhybrids.Theytendtobemorehealthy,productive,anduniformthantheirparents.

FEMALESELF

Proteinbandingpatternsthatareidenticaltotheseedparentofahybrid.

GENE

Anaturalunitofthehereditarymaterial,whichisthephysicalbasisforthetransmissionofthe

characteristicsoflivingorganismsfromonegenerationtoanother.Thebasicgeneticmaterialisfundamentallythesameinalllivingorganisms:itconsistsofchainlikemoleculesofnucleicacidsdeoxyribonucleicacid(DNA)inmostorganismsandribonucleicacid(RNA)incertainviruses andisusuallyassociatedinalineararrangementthat(inpart)constitutesachromosome.

ThesegmentofDNAthatisinvolvedinproducingapolypeptidechain.Itincludesregionsprecedingandfollowingthecodingregion(leaderandtrailer)aswellasinterveningsequences(introns)betweenindividualcodingsegments(exons).

GENETIC CODE

ThesetoftripletcodewordsinDNAcodingforalloftheaminoacids.Therearemorethan20different

amino

acids

and

only

four

bases

(adenine,

thymine,

cytosine,

and

guanine).

The

mRNA

code

is

a

triplet

code,thatis,eachsuccessive"frame"ofthreenucleotides(sometimescalledacodon)ofthemRNAcorrespondstooneaminoacidoftheprotein.Thisruleofcorrespondenceisthegeneticcode.Thegeneticcodeconsistsof64entries the64tripletspossiblewhentherearefourpossiblenucleotides,eachofwhichcanbeatanyofthreeplaces(4x4x4=64).Atripletcodewasrequiredbecauseadoubletcodewouldhaveonlybeenabletocodefor(4x4=16)sixteenaminoacids.Atripletcodeallowsforthecodingof64theoreticalaminoacids.Sinceonlyalittleover20exist,thereissomeredundancyinthesystem.Hencesomecertainaminoacidsatecodedforbytwoorthreedifferenttriplets.

GENETICS

The

branch

of

biology

concerned

with

heredity,

it

was

literally

invented

by

Gregor

Mendel

in

the

19th

century.Itisastudyofthemannerinwhichgenesoperateandaretransmittedfromparentstooffspring.Itinvolvesthestudyofthemechanismofgeneaction themannerinwhichthegeneticmaterial(DNA)affectsphysiologicalreactionswithinthecell.

GENOME

Onecompletesetofgeneticinformationfromageneticsystem;e.g.,thesinglestrand,circularchromosomeofabacteriumisitsgenome.

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GENOMICS

Thescientificstudyofgenesandtheirroleinanorganism'sstructure,growth,health,disease(and/orresistancetodisease,etc.).

GENOTYPE

The

total

genetic,

or

hereditary,

constitution

that

an

individual

receives

from

its

parents.

An

individual

organism'sgenotypeisdistinguishedfromitsphenotype,whichisitsappearanceorobservablecharacter.

GMO

Geneticallymanipulatedorganism,orgeneticallymodifiedorganism.

GOODLABORATORYPRACTICES (GLP)

AsetofrulesandregulationsissuedbytheFoodandDrugAdministration(FDA)thatestablishesbroadmethodologicalguidelinesforproceduresandrecordkeeping.Theyaretobefollowedinlaboratoriesinvolvedinthetestingand/orpreparationofpharmaceuticals.GLPsalsoapplytotheEnvironmental

ProtectionAgency(EPA)(e.g.,toxicitytestingofnewherbicides).

HELIX

Aspiral,staircaselikestructurewitharepeatingpatterndescribedbytwosimultaneousoperations(rotationandtranslation).Itisoneofthenaturalconformationsexhibitedbybiologicalpolymers.

HERBICIDETOLERANTCROP

Cropplants,cultivatedbyman,whichhavebeenalteredtobeabletosurviveapplication(s)ofoneormoreherbicidesbytheincorporationofcertaingene(s),viaeithergeneticengineeringortraditionalbreedingtechniques.Forexample,crops(e.g.,soybean,canola,cotton,corn/maize,etc.)aremadetoleranttoglyphosatecontainingherbicidesbyinsertion(viageneticengineeringtechniques)ofthe

transgeneforCP4EPSPS.Corn(maize)ismadetoleranttoimidazolinonecontainingherbicidesbyadding(viatraditionalbreedingtechniques)theimidazolinonetoleranttrait.ThattraitisimpartedbytheTGene,ITGene,ortheIRGene.

HETEROTROPH

Anorganismthatobtainsnourishmentfromtheingestionandbreakdownoforganicmatter.

HETEROZYGOTE

Anindividualorganismwithdifferentallelesatoneormoreparticularloci.

HOOKEFFECT

WhenanELISAsystemisoverwhelmedwiththetargetantigenresultinginlowerthanexpectedoptical

densityreadingsforlowerdilutionsamplesthanhigherdilutedsamples.

HORMONE

Atypeofchemicalmessenger(peptide),occurringbothinplantsandanimals,thatactstoinhibitorexcitemetabolicactivities(inthatplantoranimal)bybindingtoreceptorsonspecificcellstodeliverits

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"message."Ahormone'ssiteofproductionisdistantfromthesiteofbiologicalactivity(i.e.,wherethemessageisdelivered).

HYBRIDIZATION(MOLECULAR GENETICS)

Thepairing(tightphysicalbonding)oftwocomplementarysinglestrandsofRNAand/orDNAtogivea

double

stranded

molecule.

HYBRIDIZATION(PLANTGENETICS)

Thematingoftwoplantsfromdifferentspeciesorgeneticallyverydifferentmembersofthesamespeciestoyieldhybrids(firstfilialhybrids)possessingsomeofthecharacteristicsofeachparent.Those(hybrid)offspringtendtobemorehealthy,productive,anduniformthantheirparents aphenomenonknownas"hybridvigor".Hybridscanalsoarisefrommorethantwo("parent")species.

Hybridcom/maizeseedwasfirstcommercialized(intheUnitedStates)in1922.Otherrecentlycreatedcrophybridsincludetangelos(producedbycrossinggrapefruitwithtangerines),nectarines(bredfrompeaches),etc.

Somehybridshaveoccurredspontaneouslyinnature.Forexample,wheat(Triticumaestivum)arosecenturiesagofromanaturallyoccurringinterbreedingofthreeMiddleEastgrasses.Inthe1980s,sugarbeet(Betavulgarissubspeciesvulgaris)naturallyinterbredwiththewildnativeweedknownasseabeet(Betavulgarissubsp,maritima)inEurope;resultinginanannualweed(incontrasttosugarbeet,whichisabiannual).Becausethat(newhybridweed)iscloselyrelatedtosugarbeet,anyherbicidethatkillsthe(newhybridweed)islikelytoharmthesugarbeetcrop(unlessthesugarbeetcropismadeherbicidetolerant).

IMMUNOASSAY

Theuseofantibodiestoidentifyandquantify(measure)substancesbyavarietyofmethods.Thebindingofantibodiestoantigen(substancebeingmeasured)isoftenfollowedbytracers,suchasfluorescenceor(radioactive)radioisotopes,toenablemeasurementofthesubstance.

ISOELECTRIC POINT

ThepHatwhichaparticularmoleculeorsurfacecarriesnonetelectricalcharge.

ISOZYMES

(isoenzymes)Multipleformsofanenzymethatdifferfromeachotherintheirsubstrate(substanceactedupon)affinity,intheirmaximumactivity,orintheirregulatoryproperties.

KB

Anabbreviationfor1,000(kilo)basepairsofdeoxyribonucleicacid(DNA).

KILODALTON(KD)

Aunitofmassequalto1,000Daltons.

LIMITOF DETECTION (LOD)

Thelowestanalyteconcentrationthatcanbedetected. TheLODisnotnecessarilythelowestamountthatisquantifiedtoanexactvalue.

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LINKEDGENES

TwoormoreGenesthatareinsertedwithinthesameconstruct,intothesamechromosomelocation,andarealwayspresenttogether.IfaseedorplanthastheDNAforonegeneitalsohastheotherpresent.Itcanbeamarkergenelinkedtoageneofinterestor2ormoregenesofinterestlinkedtogether.

MALESELF

Proteinbandingpatternsthatareidenticaltothepollenparentofahybrid.

MARKER(DNAMARKER)

ADNAfragmentofknownsizeusedtocalibrateanelectrophoreticgel.

MARKER(DNASEQUENCE)

AspecificsequenceofDNAthatisvirtuallyalwaysassociatedwithaspecifiedtrait,becauseof"linkage"betweenthatDNAsequence(the"marker")andthegene(s)thatcausethatparticulartrait.

MARKER(GENETIC MARKER)Atraitthatcanbeobservedtooccurornottooccurinanorganismsuchas,forexample,bacteriaorplant(s).Geneticmarkersincludesuchtraitsas:expressionofluciferaseinleafcells(causingleavestoglow),resistancetospecificantibiotics,thenatureofthecellwallandcapsulecharacteristics,requirementsforaparticulargrowthfactor,andcarbohydrateutilization,tomentionafew.Forexample,ifacultureofdividing(growing)bacteriathatisnotresistanttoaparticularantibiotic(i.e.,lacksthetraitofantibioticresistance)isexposedtoonlytheDNAisolatedfrombacteriathatareresistanttotheantibiotic,thenafractionofthecellsexposedwilldirectlyincorporatethistrait(someDNA)intotheirgenome,henceacquiringthetrait.Thefirstgeneticallyengineeredplantsbearingamarkergenewerefieldtestedin1986.

MESSENGERRNA(MRNA)

Messengerribonucleicacid.TheintermediarymoleculebetweenDNAandribosomes(inacell)whichsynthesize(i.e.,make)thoseproteinscodedforbythecell'sDNA.Uponreceivingthe"message"encodedintheDNA,themessengerRNApassesthroughtheribosomeslikeareelofpunchedpaperpassesthroughanoldplayerpiano(pianola)givingtheribosomesthespecificationsformakingthecodedforproteins.

ThisprocessisaidedbytransferRNA(tRNA)molecules,whichforageforaminoacidsthatfloataroundinthecell(outsideofthecell'snucleusandribosomes).ThetransferRNA(tRNA)moleculesattachto,andescortindividualaminoacidstotheribosome,asandwhenthemessengerRNA(mRNA)directs.Eachofthe20differentaminoacidshasatleastoneofitsownpurposebuilttRNAmolecules,whichpossessathreelettercodeofnucleotidesatthestemofthecloverleafshapedrRNAmolecule.

TheribosomehasroomforonlytwotRNAmoleculesatatime.ThemessengerRNA(mRNA)molecule(whichitselfispassingthroughtheribosome)callsoverthefirsttRNAmolecule,whichbringswithitthespecifiedaminoacid.ShortsectionsofthemessengerRNA(mRNA)andtransferRNA(tRNA)moleculeslocktogetherinsidetheribosome(becausewherethesetwomoleculesmeet,theirthreenucleotidesarecomplementary),thewhole(lockedtogether)apparatusshiftsalongbythreenotches(i.e.,nucleotides),andasecondtRNAmolecule(bearinganotheraminoacid)slipsinnexttothefirsttRNAmolecule.

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compound.Ananalyticalinstrumentknownasaspectrophotometerisusedto(quantitatively)expresstheamountofasubstance(dissolved)inasolution.Mathematically,thisisaccomplishedusingtheBeerLambertLaw.

ORGANIZATION FORECONOMIC COOPERATION ANDDEVELOPMENT(OECD)

An

international

organization

comprised

of

the

world's

wealthiest

(most

developed)

nations.

In

1991,

theOECD'sGroupofNationalExpertsonSafetyinBiotechnology(GNE)completedadocumententitledReportontheConceptsandPrinciplesUnderpinningSafetyEvaluationsofFoodDerivedfromModernBiotechnology.The"aimofthatdocumentwastoelaboratethescientificprinciplestobeconsidered(i.e.,byOECDmembernations'regulatoryagencies)inevaluatingthesafetyofnewfoodsandfoodcomponents"(e.g.,geneticallymodifiedsoybeans,corn/maize,potatoes,etc.)

OUTCROSSING

Thetransferofagivengeneorgenes(e.g.,onesynthesizedbymanandinsertedintoaplantviageneticengineering)fromadomesticatedorganism(e.g.,cropplant)towildtype(relativeofplant). Incropproduction,outcrossingispollinationfromanundesiredpollensource

PAT GENE

Adominantgenewhich,wheninsertedintoaplant'sgenome,impartsresistancetoglufosinateammoniumcontainingherbicides.Becausetheglufosinate ammoniumherbicidesactviainhibitionofglutaminesynthetase(anenzymethatcatalyzesthesynthesisofglutamine),thisinhibitionofenzymekillsplants(e.g.,weeds).Thatisbecauseglutamineiscrucialforplantstosynthesizecriticallyneededaminoacids.ThePATgeneisoftenusedbygeneticengineersasamarkergene.

PHENOTYPICMARKER

Theoutwardphysicalappearanceofaparticulartrait.

PHYSIOLOGYThebranchofbiologydealingwiththestudyofthefunctioningoflivingthings.Thematerialsofphysiologyincludealllife:animals,plants,microorganisms,andviruses.

PlantVarietyProtectionAct(PVP)AlawpassedbytheUnitedStatesCongressin1970thatenablesintellectualpropertyprotection(analogoustopatentprotection)fornewseedplantsandseedsinAmerica.

POLYACRYLAMIDEGELELECTROPHORESIS (PAGE)

Aformofchromatographyinwhichmoleculesareseparatedonthebasisofsizeandcharge.Thestationaryphase(thepolyacrylamidegel)isapolymerizedversionofacrylamidemonomers.Thegel

looks

and

feels

like

JelloTM.

On

a

molecular

basis

it

consists

of

an

intertwined

and

cross

linked

mesh

of

polyacrylamidestrings.Ascanbeimaged,thereareholesinthegel(likeinaplasticmeshbag)andwithenoughcrosslinkingthesizeoftheholesbeginstoapproachthesizeofthemoleculeswhicharetobeseparated.Sincesomemoleculeswillbelargerandsomesmaller,someofthemwillbeabletopassthroughthegelmatrixmoreeasilythanothers.Thisispartofthebasisforseparation.

Itshouldbenotedatthispointthatifthegeliscrosslinkedenoughandbecauseofthistheholesinthatgelaresmallerthanthemoleculestobeseparated,thenthemoleculeswillnotbeabletopenetrate

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intothegelandnoseparationcanoccur.Thechargeonthemoleculealsoplaysaroleintheseparation.Functionally,thegelservestoholdandseparatethemolecules.Althoughdetailsarenotpresentedhere,afterthegelhasbeenprepared(pouredandcross linked)asmallamountofthesolutioncontainingthemoleculestobeseparatedisplacedintowells(groovestoholdtheliquid)onthegelandthesystemissubjectedtoanelectriccurrent.Overthecourseofminutestohoursmoleculesbearingdifferentcharge/massseparate.

POLYCLONALANTIBODIES

Amixtureofantibodymolecules(thatarespecificforagivenantigen)thathasbeenpurifiedfromanimmunized(tothatgivenantigen)animal'sblood.Suchantibodiesarepolyclonalinthattheyaretheproductsofmanydifferentpopulationsofantibodyproducingcells(withintheanimal'sbody).Hencetheydiffersomewhatintheirprecisespecificityandaffinityfortheantigen.Yearsago,antibodies(thencalledantitoxin)thatwerepurifiedfromanimmunizedanimal'sblood(e.g.,ahorse)wereinjectedintohumanssufferingfromcertaindiseases(e.g.,diphtheria).Inthesecasesthepathogenhadcauseddiseasebysecretinglargeamountsoftoxinintothevictim'sbloodstream.Theantitoxincombinedquantitatively(e.g.,1:1,2:1,1:2,1:3,3:1,etc.)with,andneutralizedthetoxin(forthosefewdiseasesforwhichitwasapplicable).Vaccinesarenowusedinstead,becauseoftheadverseimmuneresponsecausedbythehorse'sblood(antigens).

POLYMERASECHAINREACTION (PCR)

AreactionthatusestheenzymeDNApolymerasetocatalyzetheformationofmoreDNAstrandsfromanoriginalonebytheexecutionofrepeatedcyclesofDNAsynthesis.Functionally,thisisaccomplishedbyheatingandmeltingdoublestranded(hydrogenbonded)DNAintosinglestranded(nonhydrogenbonded)DNAandproducinganoligonucleotideprimercomplementarytoeachDNAstrand.TheprimersbindtotheDNAandmarkitinsuchawaythattheadditionofDNApolymeraseanddeoxynucleosidetriphosphatescauseanewstrandofDNAtoformwhichiscomplementarytothetargetsectionofDNA.Theprocessdescribedpreviouslyisrepeated(trait,product,etc.)againandagaintoproducemillionsofcopiesofthedesiredstrandofDNA.PCRanditsregisteredtrademarksarethepropertyofF.Hoffmann

LaRoche&Co.AG,Basel,Switzerland.

POLYMERASECHAINREACTION (PCR)TECHNIQUE

Developedin1984and1985byKaryB.Mullis,RandallK.Saiki,StephenJ.Scharf,FredA.Faloona,GlennHorn,HenryA.Erlich,andNormanArnheim,thePCRtechniqueisaninvitromethodthatgreatlyamplifies(makesmillionsofcopiesof)DNAsequencesthatotherwisecouldnotbedetectedorstudied.ItcanbeutilizedtoamplifyagivenDNAsequencethatconstituteslessthanonepartpermillionofinitialsample(e.g.,a100basepairtargetDNAsequencewithinthegenomeofoneofthehigherorganisms,whichcancontainupto500millionbasepairs),TheprocedurealleviatesthenecessityofinvivoreplicationofatargetDNAsequence,orofreplicationofoneofakindtinyDNAsamples(e.g.,fromacrimescene).

PRECISION

Measuredbyrepeatabilityofresults,oneofthefourcriteriaofmethodvalidationforpuritytesting.

PRIMER(DNA)

Ashortsequencedeoxyribonucleicacid(DNA)thatispairedwithonestrandofthetemplateDNA.ItisthegrowingendoftheDNAchainanditsimplyprovidesafree3'OHendatwhichtheenzymeDNA

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polymeraseaddsondeoxyribonucleotideunits(monomers).WhichdeoxyribonucleotideisaddedisdictatedbybasepairingtothetemplateDNAchain.WithoutaDNAprimersequenceanewDNAchaincannotformsinceDNApolymeraseisnotabletoinitiateDNAchains.

PROMOTER

The

region

on

DNA

to

which

RNA

polymerase

binds

and

initiates

transcription.

The

promoter

"promotes"thetranscription(expression)ofthatgene.AregionofDNA(deoxyribonucleicacid)whichlies"upstream"ofthetranscriptionalinitiationsiteofagene.Thepromotercontrolswhere(e.g.,whichportionofaplant,whichorganwithinananimal,etc.)andwhen(e.g.,whichstageinthelifetimeofanorganism)thatthegeneisexpressed.Forexample,thepromoternamed"Bce4"is"seedspecific"[i.e.,itonly"promotes"theexpressionofagivengene'sproduct(e.g.,protein,fattyacid,aminoacids,etc.)withinaplant'sseed].

PROTEIN

FromtheGreekwordproteios,whichmeans"thefirst"or"themostimportant."Anyofaclassofhighmolecularweightpolymercompoundscomposedofavarietyofaminoacidsjoinedbypeptidelinkages.Viathesynthesis(ofthis"chain")performedbyribosomes,eachproteinistheultimateexpressionproductofagene.Morethanoneproteincanbeexpressedfromagivengene(theparticularproteinexpressedisdeterminedbyfactorssuchasthecell'stemperatureorotherenvironmentalvariable,presenceofSTATs someofwhichthemselvesareproteins,etc.).

Duringtheirsynthesis(afteremergingfromcell'sribosome),proteinsmayalsobephosphorylated(i.e.,a"phosphategroup"isaddedtotheproteinmolecule),glycosylated(i.e.,oneormoreoligosaccharidesisaddedontotheproteinmolecule),acetylated(i.e.,oneormore"acetylgroups"isaddedtotheproteinmolecule),farnesylated(i.e.,a"farnesylgroup"isaddedtotheproteinmolecule),ubiquinated(i.e.,aubiquitin"tag"isaddedtotheproteinmolecule),sulfated(i.e.,a"sulfategroup"isaddedtotheproteinmolecule),orotherwisechemicallymodified.Proteinsarethe"workhorses"oflivingsystemsandincludeenzymes,antibodies,receptors,peptidehormones,etc.Proteinsinlivingorganismsrespondto

changingenvironmentalandotherconditionsbychangingtheirlocationwithincells,bygettingcutinto(specific)pieces,bychangingwhich(other)moleculestheywillbind(adhere)to,etc.Alloftheaminoacidscommonlyfoundin(eachandeveryoneofthe)proteinshaveanasymmetriccarbonatom,excepttheaminoacidglycine.Thustheproteinispotentiallychiralinnature.

RANDOMAMPLIFIEDPOLYMORPHICDNA(RAPD)TECHNIQUE

AgeneticmappingmethodologythatutilizesasitsbasisthefactthatspecificDNAsequences(polymorphicDNA)are"repeated"(i.e.,appearinsequence)withgeneofinterest.Thus,thepolymorphicDNAsequencesarelinkedtothatspecificgene.Theirlinkedpresenceservestofacilitategeneticmapping(i.e.,"location"ofspecificgene(s)onanorganism'sgenome).

RECESSIVEALLELE

DiscoveredbyGregorMendelinthe1860s,thisreferstoanallelicgenewhoseexistenceisobscuredinthephenotypeofaheterozygotebythedominantallele.Inaheterozygotetherecessivealleledoesnotproduceapolypeptide;itisswitchedoff.Inthiscasethedominantalleleistheoneproducingthepolypeptidechain.

RECOMBINANT DNA(RDNA)

DNAformedbythejoiningofgenes(geneticmaterial)intoanewcombination.

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RENATURATION

Thereturntothenaturalstructureofaproteinornucleicacidfromadenatured(morerandomcoil)state.Forexample,aproteinmaybedenatured[loseitsnative(natural)structure]byexposuretosurfactantssuchasSDSortochangesinthepHofthemedium,etc.IfthesurfactantisslowlyremovedorthepHisslowlyreadjustedtotheoptimumfortheprotein,itwillrefold(snap)backintoitsoriginal

(native)

form.

RIBONUELEIC ACID(RNA)

Alongchain,usuallysinglestrandednucleicacidconsistingofrepeatingnucleotideunitscontainingfourkindsofheterocyclic,organicbases:adenine,cytosine,guanine,anduracil.Thesebasesareconjugatedtothepentosesugarriboseandheldinsequencebyphosphodiester(chemical)bonds.TheprimaryfunctionofRNAisproteinsynthesiswithinacell.However,RNAisinvolvedinvariouswaysintheprocessesofexpressionandrepressionofhereditaryinformation.ThethreemainfunctionallydistinctvarietiesofRNAmoleculesare:(1)messengerRNA(mRNA)whichisinvolvedinthetransmissionofDNAinformation,(2)ribosomalRNA(rRNA)whichmakesupthephysicalmachineryofthesyntheticprocess,and(3)transferRNA(tRNA)whichalsoconstitutesanotherfunctionalpartofthemachineryofproteinsynthesis.

RUGGEDNESS

Reproducibilityofresultsobtainedundervaryingconditionssuchasdifferentlabsandequipment.

SDS

Sodiumdodecylsulfate.Alsoknownassodiumlaurylsulfate(SLS).Asurfactantcommonlyusedinbiochemicalandbiotechnologicalapplicationsforthesolubilizationofmembranecomponentsandhardto solubilize(dissolve)molecules.Forexample,itisoftenutilizedathighconcentrationinwatersolution(e.g.,alongwithpotassiumacetate)todissolveplantDNAsamples(e.g.,whenascientistwantstosequencethatsampleofplantDNA).TheSDS/PAinwatersolutionhelpsthescientisttoseparateout

contaminants

that

are

commonly

present

in

samples

from

plant

tissues

(i.e.,

polysaccharides,

proteins,

etc.)becauseDNAmoleculesaremuchmoresolubleinSDS/PAsolutionthanarethosecontaminantmolecules.Aboveacriticalconcentration(CMC),SDSformsmicellesinwaterwhicharethoughttoberesponsibleforitssolubilizingaction.SDSisalsousedinsuchitemsasshampoo.

SEGREGATION

SegregationistheFirstLawofMendeliangenetics. Gametesfromanyheterozygousparentseparateandpropagateinamannerthatisindependentoftheallelesofthesamegeneandalltheothergenesinthegenome. Becauseofthis,geneticpuritycanbedeterminedforanygivengene(TraitPurity)oranygroupofgenes(HybridorVarietalPurity)

SELF

Apistolthatisfertilizedfrompollenfromthesameplantthatbearsthepistil,thetermalsoreferstotheseedresultingfromsuchfertilizations.

SEQUENCE(OFADNAMOLECULE)

ThespecificnucleicacidsthatcompriseagivensegmentofaDNAmolecule.

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SEQUENCE(OFAPROTEINMOLECULE)

Thespecificaminoacids(andtheorderinwhichtheyarecoupledtogether)thatcompriseagivensegmentofaproteinmolecule.

SEQUENCING (OFDNAMOLECULES)

The

process

used

to

obtain

the

sequential

arrangement

of

nucleotides

in

the

DNA

backbone.

The

cleavageintofragments(followedbyseparationofthosefragments,whichcanthenbesequencedindividually)ofDNAmoleculesbyoneofseveralmethods:(1)achemicalcleavagemethodfollowedbypolyacrylamidegelelectrophoresis(PAGE),(2)amethodconsistingofcontrolledinterruptionofenzymaticreplicationmethodsfollowedbyPAGE,(3)adidexylmethodutilizingfluorescent"tag"atomsattachedtotheDNAfragments,followedbyuseofspectrophotometrytoidentifytherespectiveDNAfragmentsbytheirdiffering"tags"(whichfluoresceatdifferentwavelengths).This(fluorescenttag)variantofthedideoxymethodcanbeautomatedto"decipher"largeDNAmolecules(i.e.,genomes).Suchautomatedmachinesaresometimescalled"genemachines."

"SHOTGUN" METHOD

[to

introduce

foreign

(new)

genes

into

plant

cells]

A

technique

for

gene

into

cell

introduction

in

which

thegeneisattachedtotiny"bullets"madeoftungstenorothermetal.Bymeansofaspecialdevice("genegun")thetinyparticlesarethenliterally"shot"throughtheplasmamembraneintoplantcellswith:

(a)Highpressuregas(e.g.,theGENEBOOSTERgundevelopedatHungary'sAgriculturalBiotechnologyCenterutilizesnitrogen).

(b)Aratherconventionalfirearm(sometimescalledaparticlegun)whichusesa.22calibershellminustheleadtip.Thetinyparticlesareusedinplaceoftheleadtip.Forexample,theBIOLISTICGeneGuninventedatAmerica'sCornellUniversityutilizes"bullets"madeoftungsten.

Someplantcellsaredestroyedintheprocessandthesurvivorsheal(providedthe"bullet"issmallenough),andincorporate(some)ofthenewgeneticmaterialintotheirgeneticcomplement,andproduceswhateverproduct(i.e.,aprotein)thenewlyintroducedgenecodesfor.

SPECIFICITY

Theabilitytoaccuratelymeasureatargetanalyteinthepresenceofothercomponentsthatmaybepresent. Typicallythesemightincludeproteins,nucleicacids,impurities,degradants,matrixandbuffers

"STACKED"GENES

Referstotheindependentinsertionoftwoormore(synthetic)genesintothegenomeofanorganism.OneexampleofthatwouldbeaplantintowhichhasbeeninsertedagenefromBacillusthuringiensis(B.t.)andageneforresistancetoaspecificherbicide.

SUBSTRATE(CHEMICAL)

Thesubstanceactedupon,forexample,byanenzyme.Forexample,theenzymeamylasebreaksstarchdownintoglucosemolecules;starchisthesubstrate(oftheenzymeamylase).

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