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1、禽病原性大腸桿菌E058株ompT、cvaC基因突變株的構(gòu)建及其生物學(xué)特性ConstructionandcharacterizationofompTandcvaCmutantsotavlanPathogemc15scherichiacoli^●1●1—1·11‘本文受國家自然科學(xué)基金(30972196、30771604、30471281)、國家863計劃(2003AA222141)資助研究生:何洪波專業(yè):預(yù)防獸醫(yī)學(xué)導(dǎo)師:高崧教授劉秀梵教授揚州大學(xué)獸醫(yī)學(xué)院2012年6月何洪波:禽病原性人腸桿菌E058株ompT、n,臼(1基閃突變株的構(gòu)建及其生物學(xué)特性上禽病原性大腸桿菌E058株ompT、cva
2、C基因突變株的構(gòu)建及其生物學(xué)特性;開要三、由㈣y225㈣8803研究生:何洪波揚州入學(xué)碩十學(xué)位論文生株相比沒有明顯的差異,從而表明ompT基因與APECE058株的致病性有關(guān),cvaC與APECE058株的致病性不相關(guān)。關(guān)鍵詞:禽致病性大腸桿菌;ompT:cvaC;突變株;致病性何洪波:禽病原性人腸桿菌E058株ompT、cva(1基岡突變株的構(gòu)建及其生物學(xué)特性三ConstructionandcharacterizationofompTandcvaCmutantsofaVlanpamogenlcEsc刃ericiqlacoli’1·11T‘1·1‘AbstractM.S.candidate:H
3、eHongboAdvisor:Prof.GaoSongProf.LiuXiufanColibacillosisreferstoanylocalizedorsystemicinfectioncausedentirelyorpartlybyavianpathogenicEscherichiacoli(APEC),includingsepticaemia,granuloma(Hjarre’Sdisease),synovitis,airsacdisease/chronicrespiratorydisease(CRD),peritonitis,salpingitis,aviancellulites,sw
4、ollenheadsyndrome,panophthalmitis,andomphalitis/yolksacinfection.Collectively,infectionscausedbyEscherichiacoli(Ecoli)areresponsibleforsignificanteconomiclossestothepoultryindustryinmanypartsoftheworld.ompTencodesOmpTprotease.a(chǎn)ndcvaCisthestructuralgeneforcolicinVEpidemiologicsurveyshowsthatompTandcv
5、aCareofhighratiointhevirulencestrains.Inthisstudy,therelationshipbetweenompTgeneorcvaCgeneandthepathogenicityofAPECstrainE058wasstudied.AlltheactualrolesoftheknownvirulencefactorsofAPECarenotfullyelucidatedandcertainstepsoftheinfectionprocesshavenotbeenrelatedtopreviouslyidentifiedfactors.Furthermor
6、e,themechanismsbywhichAPECcauseinfectionarelargelyunknown.Inthisstudy,genesinAPECserogroup02strainE058chosenfordeletionwereompTandcvaC.ThefragmentswereamplifiedandsubclonedtopMD⑩18.Tsimplevector.ThevectorT-ompTwasinsertedwithaZeo。genetoformT-ompT-ZeoandT-cvaCwasinsertedwithaZeo。genetoformT-cvaC-Zeo.
7、ThedisruptedompTandcvaCgeneswithZeo。geneinsertedinT-ompT-ZeoandT-cvaC—ZeowereamplifiedbyPCRandpurifiedforelectroporationtoE058competentcells,theresultingZeo‘colonieswereselectedandnamedasE058AompT,E05