資源描述:
《黑曲霉β甘露聚糖酶基因的克隆與其原核和真核表達》由會員上傳分享,免費在線閱讀,更多相關內容在學術論文-天天文庫。
1、AbstractAspergillusniger13-mannanasegenecloningandexpressionofprokaryoticandeukaryotict;-mannanase(endo-1,4-13·D-mannanase;mannohydrolase;EC3.2.1.78)areakindofclassabletohydrolyzeoligosaccharides,mannosepolysaccharides(includingmannart,galactomammn,glu
2、comannan,etc.)of1,4-/3一D—mannosidaseendobondhydrolaseandasemi-celluloseenzymes.Theapplicationprospectof13-mannanaseisveryextensivestudyofitshi;ghcommercialvalueandscientificsignificance.Now13-mannanasehasbeenwidelyusedinthefood,pharmaceutical,papermaki
3、ng,textilepriminganddyeing,oilexplorationandotheraspectsofbiologicalresearchtechniques.IndustrialproductionofAspergillusnigeristhemainspeciesmatlnanase.However,thecurrentproductionstrainexistsalargenumberofdisadvantages,suchasanumberofsporesduringferme
4、ntation,shortfermentationperiod,lowenzymeproduction.Inthiscase,theapplicationofconstructionofgeneticengineeringbacteriamannanase,simplifiedfermentationconditions,increasedenzymeproductioIl,haveimportantresearchvalue.Inthisresearch,mannanasegeneproducin
5、gstrainofAspe哂llusnigerasagenedonor,clonedmanrlanasegene(MAN),wereseperatelyconstructedtheE.coliexpressionvector,PichiapastorisexpressionvectorandtheAgrobacterium-mediatedgeneMANhomologousrecombinationvector,respectively,inthemeantime,weretransferredto
6、E.coliBL21,PichiapastorisGS15andglucoamylaseproducingstrainofAspergillusnigerforlayingthefoundationforhi曲lyexpressedgenes-Themainresultsare私follows:Itapplied13-mannanaseproducingstrainofAspergillusnigerasthematerialandobtainedbyRT-PCRamplificationtogai
7、nlB-marmanasegeneMANoftheeDNAfragments.ThoughPCRamplificationofIB-mannanasegenegainedMANoftheDNAfragments,thematerialthatisAspergillusnigerglucoamylaseproductionobtainedbyPCRamplificationGIAgene5。and3’homologousarmoftheDNAfragments.Asaresult,thesequenc
8、eiscorrect.1.MANgeneexpressioninE.coil.Itapplied13-mannanaseproducingstrainofAspergillasnigerasthematerialandobtainedbyRT-PCRamplificationtogainlB-mannanasegeneMANoftheeDNAfragments.MANgeneore.coilwasconstructedtoexpressvectorpET-MAN。Af