Rgt Study Guide 4-1-2010
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Transcript of 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
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
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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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