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1、江南大學(xué)碩士學(xué)位論文Ⅱa類(lèi)乳酸菌細(xì)菌素的克隆表達(dá)研究姓名：李碩申請(qǐng)學(xué)位級(jí)別：碩士專(zhuān)業(yè)：糧食、油脂及植物蛋白工程指導(dǎo)教師：張灝20090301AbstractAbstractBacteriocinsproducedbylacticacidbacteria(LAB)areaheterogeneousgroupofactivatedpeptideorproteinousmaterial，mostofwhichareprovidedwithin

2、hibitionforbacteriainnearrelationandtheminoritywithwiderbacteriostaticspectraBacteriocinscanbedividedintofourtypesClassIIabacteriocinsCanbeconsideredasthemajorsubgroupofbacteriocinsfromlacticacidbacterianotonlybecauseoft

3、heirlargenumber，butalsobecauseoftheiractivitiesandpotentialapplicationsTheyhavefirstattractedparticularattentionaslisteriamonocytogenescompoundsandarenowbelievedtobethenextinlineifmorebacteriocinsaretobeapprovedinthefutu

4、reBecauseofhisantilisterialeffectiveness，classIIabacteriocinshavesignificantpotentialasbiopreservativesinalargenumberoffoodsEachoflacticacidbacteriaproducesatleastoneormoreofthebacteriacinAndSOfaronly29classIIaofbacteria

5、cinswerefound，mostofwhichwerediscoveredinthe1990sIsolationandpurificationoftheproteinfromnativehostspurificationaretimeconsuming，laboriousprocesses，whichusuallyresultinlowproteinyieldSoitismuchdifficulttofindanewclassIIa

6、bacteriocinnowButwecancontinuetostudytheclassIIabacteriocinsbybioinformaticsandmolecularbiologymeathodSakacinP，whichisaclassIIabacteriocin，isveryactiveagainstthefoodpathogenListeriamonocytogenesPediocinPA一1isoneofIIabact

7、edocinswhichhavebeenusedasfoodpreservativeInthisstudytheclonevectors(pGMTsakPpGMTped)，expressionvectors(pET28asakPpET28aped)ofPediocinPA一1andSakacinPwereconstructed，thenthetargetproteinsweresuccessfullyexpressedinEcoliBL

8、21(DE3)andtheirantibacterialactivitywerealsoobtainedAndwealsofoundthatwiththereductionofincubationtemperature，theexpressionlevelofsolubleproteinincreasesdramaticallyTheseresultswillbehelpfulforestablishingagoodmethodforn

9、ewIIabacteriocinscreeningBybioinformaticanddatabasesearching，asegmentofgeneswhichhashighsequencesimilaritieswiththematurepeptideofclassIIabacteriocinwasfoundoutWenameditNBC1，anditsfunctionisnotidentifiedyetFirstly，wesynt

10、hesizedthegenesequenceofNB—C1artificially，andconstructedtheclonevector(pGMTC1)，expressionvector(pET28aC1)ThenthetargetproteinwassuccessfullyexpressedinEcoliBL21(DE3)anditsantibacterialactivityagainstlisteriamonocytogenes

11、WasalsoobtainedTherefore，wecanconcludethatNB—C1isanewclassIIabacteriacinproducedbyLABThesuccessfulcloningandexpressionofbiologicallyactivesakacinPandNB—C1inEcoliBL21(DE3)maybehelpfultodeterminemorenovelclassIIabacterioci

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