基因工程课后题英文版.docx
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基因工程课后题英文版.docx
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基因工程课后题英文版
第一章为什么基因克隆和DNA分析很重要
geneticengineering:
isthedirectmanipulationofanorganism’sgenomeusingmodernDNA
technology.ItinvolvestheintroductionofforeignDNAorsyntheticdenesintotheorganismofinterest.
1.HowtoCreateaGeneticallyModifiedPlant?
Ans:
①Createrecombinantbacteriawithdesiredgene.
②Allowthebacteriato“infect”theplantcells.
③Desiredgeneisinterestedintoplantchromosomes.
2.WhygenecloningandPCRaresoimpartant?
Ans:
becausebothtechniquescanprovideapuresampleofanindividualgene,separatedfromalltheothergenesinthecell.
第二章基因克隆的载体:
质粒和噬菌体
1.Pleaselistessentialpartsofacloningvetor.
(1)Self-replication:
orisite.
(2)Plasmidsize:
assmallaspossible.
(3)Insertsite:
MCS(Multi-cloningsite).
(4)Selectablemarker.
(5)Notaffecthostcell.
2.Classificationofplasmid.
1)Basedonthemaincharacteristiccodedbytheplasmidgene.
①FertilityorFplasmids
②ResistanceorRplasmids
③Colplasmid
④Degradativeplasmids
⑤virulenceplasmids
2)dependonwhethertheycarrythetragenes:
①conjugativeplasmids
②non-conjugativeplasmids.
3)basedonthecopiestheycarrying:
relaxedplasmids;stringentplasmids
Words:
1.Plasmid:
AusuallycircularpieceofDNA,primarilyindependentofthehostchromosome,oftenfoundinbacteriaandsomeothertypesofcells.
2.cccDNA:
(covalentlyclosed-circularDNA):
Acompletelydouble-standedcircularDNAmolecule,withnonicksordisconuities,usuallywithasupercoiledconformation.
3.ori(originofreplication:
ThespecificpositiononaDNAmoleculewhereDNAreplicationbegins.
4.episome:
Aplasmidcapableofintegrationintothehostcell'schromosome.
5.copynumber:
Thenumberofmoleculesofaplasmidcontainedinasinglecell.
6.bacteriophage:
Aviruswhosehostisabacterium。
BacteriophageDNAmoleculesareoftenusedascloningvectors。
7.2μmplasmid:
AplasmidfoundintheyeastSaccharomycesCerevisiaeandusedasthebasisforaseriesofcloningvectors.
8.cossite:
thebacteriophagecohesiveendsarecalledcossite。
9.Pilus:
oneofthestructurespresentonthesurfaceofabacteriumcontainingaconjugativeplasmid,throughwhichDNAisassumedtopassduringconjugation.
第3章从活细胞中纯化DNA
1.Whichstepsisincludedintheplasmidextractionfrombacterial?
Pleaselistthemainchemical
componentsanddescribetheirfunctionsinplasmidextractions.
①Acultureofbacteriaisgrownandthenharvested.
②Thecellsarebrokenopentoreleasetheircontents.
③ThiscellsextractistreatedtoremoveallcomponentsexcepttheDNA.
④TheresultingDNAsolutionisconcentrated.
Lysozyme:
digestthepolymericincellwall.。
EDTA:
removemagnesiumionsandinhibitenzymesthatdegradeDNA.
SDS:
removelipidmoleculesinthecellmembranes.
Removeproteinfromcellextractphenol(苯酚)or1:
1mistureofphenoland
chloroform(氯仿);
Whenproteincontentisgreatincellextract:
treatwithaprotease
RNAintheextractwasmovedbyribonuclease
CollectingDNAbyethanolprecipitation
2.Techniquesforbreakingbacterialcells.
Physicalmethods:
ultrasonic
Chemicalmethods:
lysozyme/EDTA/SDS
3.whatisthemostfrequentlyusedmethodofconcentration?
P32Ethanolprecipitation
5.Pleasedescribethegenomeextractionprocedurefromyeast.①Acultureofbacteriaisgrownandthenharvested.
②Thecellsarebrokenopentoreleasetheircontents.
③ThiscellextractistreatedtoremoveallcomponentsexcepttheDNA.
④TheresultingDNAsolutionisconcentrated。
1.Brothculture:
Theculturemediummustprovideabalancedmixtureoftheessentialnutrientsatconcentrationsthatwillallowthebacteriatogrowanddivideefficiently.
2.alkalinedenaturation:
thebasisofthistechniqueisthatthereisanarrowPh.rangeatwhichnonsupercoiledDNAisdenatured,whereassupercoiledplasmidarenot.
3.densitygradientcentrifugation:
Separationofmoleculesandparticlesonthebasisofbuoyantdensity,bycentrifugationinaconcentratedsucroseorcesiumchloridesolution.
4.plasmidamplification:
Amethodinvolvingincubationwithaninhibitorofproteinsynthesisaimedatincreasingthecopynumberofcertaintypesofplasmidinabacterialculture.
第4章DNA纯化后的利用
1.whenpuresamplesofDNAareprepared,howdowedotheminthenextstep?
ANS:
constructionoftherecombinantDNAmolecule.
2.PleaselistclassificationofDNAmanipulativeenzymes.
①nucleases②ligases③polymerases
④DNAmodifyingenzymes⑤topoisomerases
3.PleaselistfourtypesofDNApolymerase,anddescribetheirfunction.
①DNApolymeraseI:
polymerizationanddegradationDNA.
②Klenowfragment:
polymerizationDNAbutnotdegradation.
③TaqDNApolymerase:
resistanttodenaturationbyheattreatment.
④Reversetranscriptase:
SynthesisofcomplementaryDNAstrandsbyasingleRNAtemplate.
4.whatisthecentralfeatureoftypeIIrestrictionendonucleases.
ANS:
eachenzymehasaspecificrecognitionsequenceatwhichitcutsaDNAmolecule.
5.puttingstickyendsontoablunt-endedmolecule.
①Linkers②Adaptors③Homopolymertailing
6.howtoincreasetheefficiencyofligation?
①Theligationofcomplementarystickyendsincreasestheefficiencyofligation.
②Increasetheperiodoftimethattheendsareincontactwithoneanother.
③BluntendligationshoudbeperformedathightDNAconcentrations.
1.nucleases:
nucleasesareenzymesthatcut,shorten,ordegradednucleicacidmolecules.
2.ligases:
joinnucleicacidmoleculestogether.
3.DNApolymerases:
areenzymethatsynthesizesanewstandofDNAcomplementarytoanexistingDNAorRNAtemplate.
4.Exonucleases:
removenucleotidesoneatatimefromtheendofaDNAmolecule.
5.Endonucleases:
areabletobreakinternalphosphodiesterbondswithinaDNA
molecule.
6.Alkalinephosphatase(碱性磷酸酶)whichremovesthephosphategrouppresentatthephosphategrouppresentat5’terminusofaDNAmolecule.
7.Polynucleotidekinase(多核苷酸激酶)addingphosphategroupsontofree5'termini.
8.Terminaldeoxyribonucleotidestransferase(末端脱氧核苷酸转移酶)whichaddsoneormoredeoxyribonucleotidesontothe3'terminusofaDNAmolecule.
9.Doubledigestions:
inwhichtheDNAiscutbytworestrictionendonucleasesatonce.
10.partialdigestion:
carriedoutunderconditionsthatresultinthecleavageofonlyalimitednumber
oftherestrictionsitesonanyDNAmolecule.
11.Linkers:
areshortpiecesofdouble-strandedDNAwithbluntends,ofknownnucleotidesequence
whichcontainingasiteofarestrictionendonuclease,thataresynthesizedinthetesttube.
12.Adaptors:
areshortsyntheticoligonucleotideswithonestickyendofarestrictionendonuclease.
第5章将DNA引入活细胞
1.pleaseexplainthemainpurposesofcloning?
①Amplification,itallowsalargenumberofrecombinantDNAmoleculestobeproducedfromalimitedamountofstartingmaterial.
②Purification.。
2.Whataretheligationproduction?
1)ThedesiredrecombinantDNAmolecule.
2)Unligatedvectormolecules.
3)UnligatedDNAfragments.
4)Self-ligatedvector.
5)WrongrecombinantDNAmolecules.
3.Pleaselistthemethodsofidentifyingrecombinants.
①Insertionalinactivation.pBR322
②Lacselection.pUC8
4.Pleaselistthemethodsoftransformingnon-bacterialcells.
1)MethodsforintroducingnewDNAintoanimalsandplantcells.
①PrecipitationofDNAontoanimalscells.
②IntroductionofDNAintoanimalcellsbyliposomefusion.
③Transformationofplantprotoplasts.
2)TwophysicalmethodsforintroducingDNAintocells.
①Microinjection.
②Bombardingthecellswithhigh-velocitymicroprojectiles.
Words:
1.transformation:
TheintroductionofanyDNAmoleculeintoanylivingcell.
2.competentcell:
AcultureofbacteriathathasbeentreatedtoenhancetheirabilitytotakeupDNAmolecules.
3.insertionalinactivation:
AcloningstrategywherebyinsertionofanewpieceofDNAintoavectorinactivatesagenecarriedbythevector.
4.lacselection:
AmeansofidentifyingrecombinantbacteriacontainingvectorsthatcarrythelacZ'gene.Thebacteriaareplatedonamediumthatcontainsananalogueoflactosethatgivesabluecolourinthepresenceofβ-galactosidaseactivity.
5.electroporation:
AmethodforincreasingDNAuptakebyprotoplaststhroughpriorexposuretoahighvoltage,whichresultsinthetemporaryformationofsmallporesinthecellmembrane.
6.microinjection:
AmethodofintroducingnewDNAintoacellbyinjectingitdirectlyintothemolecule.
7.protoplast:
Acellfromwhichthecellwallhasbeencompletelyremoved.
第6章大肠杆菌的克隆载体
1.therequirementsforaDNAmoleculeasacloningvehicles?
①Self-replication.
②Lowmolecularweight.
③selectablemarker.
④Multiplecloningsites.
⑤Noharmfultothehostcells.
2.Propertiesofanidealcloningvehicle?
1)Easyofpurification.
2)Hightransformationefficiency.
3)Convenientselectablemarkersfortransformantsandrecombinants.
4)TheabilitytoclonereasonablylargepiecesofDNA.
3.WhyhaspBR322beensuchapopularcloningvector?
①Sizeis4363bp.
②Twosetsofantibioticresistancegenes.
③Highcopynumber.
4.pleaselistTHREEtypicalE.coliplasmidcloningvectors.
①pBR322②pBR327③pUC8orpGEM3Z
1.Cosmid:
Ancloningvectorconsistingoftheλcossiteinsertedintoaplasmid,usedtocloneDNAfragmentsupto40kbinsize.
2.Genomiclibrary:
Acollectionofclonessufficientinnumbertoincludeallthegenesofaparticularorganism.
3.Bacterialartificialchromosome(BAC):
AncloningvectorbasedontheFplasmid,usedforcloningrelativelylargefragmentsofDNAinE.coli.
4.P1-derivedartifcialchromosome(PAC):
Acloningvect
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