1、東北林業(yè)大學碩士學位論文納豆激酶基因在釀酒酵母中表達的研究姓名：孫立娜申請學位級別：碩士專業(yè)：微生物學指導(dǎo)教師：趙敏20070601東北林業(yè)大學碩士學位論文AbstractNattokinase(NK)isakindoffibrinolyticenzymethatisextractedfromnatto—atraditionalfoodinJapanComparedwithotherthrombolyticdrugswhichareus

2、edincommon，NKissaferandeasiertobeabsorbedbybody，andhasmoredirectandmorepersistenteffectonthrombosis，besidesitscostislowerThereforeitcanbedevelopedasanewandpotentfibrinolyticmedicineSaccharomycescerevisiaeisrecognizedasak

3、indofsimplesinglecelleukaryotes，whichhasnotoxinandnopathogenicityIthasbeenusedinbreadandalcoholindustryforyearsandwasaffirmedbyFDAtobeakindofbiologicalsafetyThegenomicofBacillussubticisVatnatto11086wasextractedandnattoki

4、nase(NK)genewasamplifiedbyPCRandWasIinkedandtransformednepositiveplasmidpMDl8TNKWasscreenedSequenceanalysisshowedthattheNKgeneWas1192bpinlengthincludingsignalregionpreleadingregionandmatureregionfragmentsnepurposegenehad

5、95％and69％homologywiththatofaccessionNoAY2199011fromBacillussubtilisandaccessionNoDQl786581fromDouchfbacteriarcponedinNCBI，respectivelyThehomologywasmuchdifferentItindicatedthatwecantakefurtherstudyofthedifferencesaboutna

6、ttokinasegenebetweenBacillussubtilisanddouchfbacteriawhichIcadtodifferentfunctionsTherecombinantexpressionvectorpYES2NKWastransformedintoSaccharomycescerevisiaeH158competentcellTherecombinantH158NKWasinducedwith13galacto

7、seThefibrinolyticactivityoffermentationsupematantwasdeterminedwithfibrinplateTherecombinantH158NKwasconstructedsuccessfullybringsnewideastodevelopanewroutetoconstructtheefficientgenetictransformationofSaccaromycescerevis