禽病原性大肠杆菌e058株ompt、cvac基因突变株的构建及其生物学特性

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1、禽病原性大肠杆菌E058株ompT、cvaC基因突变株的构建及其生物学特性ConstructionandcharacterizationofompTandcvaCmutantsotavlanPathogemc15scherichiacoli^●1●1—1·11‘本文受国家自然科学基金(30972196、30771604、30471281)、国家863计划(2003AA222141)资助研究生：何洪波专业：预防兽医学导师：高崧教授刘秀梵教授扬州大学兽医学院2012年6月何洪波：禽病原性人肠杆菌E058株ompT、n，臼(1基闪突变株的构建及其生物学特性上禽病原性大肠杆菌E058株ompT、cva

2、C基因突变株的构建及其生物学特性；开要三、由㈣y225㈣8803研究生：何洪波扬州入学硕十学位论文生株相比没有明显的差异，从而表明ompT基因与APECE058株的致病性有关，cvaC与APECE058株的致病性不相关。关键词：禽致病性大肠杆菌；ompT：cvaC；突变株；致病性何洪波：禽病原性人肠杆菌E058株ompT、cva(1基冈突变株的构建及其生物学特性三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．andcvaCisthestructuralgeneforcolicinVEpidemiologicsurveyshowsthatompTandcv

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