豬傳染性胸膜肺炎放線桿菌的分離鑒定及ApxⅠA、TbpB基因的克隆和原核表達研究.pdf

1、四川農(nóng)業(yè)大學碩士學位論文豬傳染性胸膜肺炎放線桿菌的分離鑒定及ApxⅠA、TbpB基因的克隆和原核表達研究姓名：余光勇申請學位級別：碩士專業(yè)：預(yù)防獸醫(yī)學指導教師：郭萬柱20080601IsolationandIdentficationofActinobcillusPleuropneumoniaeandItsApxlAandTbpBGeneCloningandProkaryoticexpressionYuGuangyong(Preventi

2、veVeterinaryScience)DirectedbyProfessorGuo繆鋤zhu(CollegeofAnimalScienceandTechologySichuanAgricultureUniversityVaan，Sichuan，China625014)AbstractOutbreakActinobcilluspleuropneumoniaefromthepiginSichuanastrainbacteriumwasis

3、olatedfromthatfarmwhichWasidentifiedasserotype薹ofActinobcilluspleuropneumoniaebymorphological，serologicalandmolecularbiologytestsTheantibioticsensitivitytestimpliedwithitWassensitivewiththetetracycline，Ceffriaxone，Norflo

4、xacin，TrimethopdmSulfamethoxazole，ciprofloxaein，anddrugresistancewiththeKanamycin，Amikacin，Cefalexin，Vancomycin，Ampicillin，GentamicinThePCRidentificationrefertOforeignreportAccordingtotheconservativeweakhaemolyticactivit

5、ygenesequences(APXlVA)ofActinobeilluspleuropneumoniaewedesignedtheprimersandidentifiedthestrainbyPCRAccordingtotheseqenceoftheApxIA(X68595)andTbpB(U16019)geneofActinobacilluspleuropleumoniae(serotype1)inGenBank，twopairof

6、thespecificprimmweredesignedandsynthesized，templated姆thegenomicⅨ姨samplesfromthestainofAPPCYYThePCRfragmentofApxIAandTbpBgenewereclonedabout3089bpand736bpThetwoamplifiedfragmentWasclonedintopMD18Tvectorandthensequencedana

7、lysisTheresultindicatesthattheclonedgeneWasacompleteORFApxIAandTbpBgene。Sequencecomparisonbetweenthedifferentserotypel，5，9，10ApxlAgeneofActinobcilluspleuropneumoniaethathomologyarebetween968988％Sequencecomparisonbetweent

8、woserotypelTbpBgeneofActinobcillus曩europneurnoniaethathomologyale997％and996％AccordingtoenzymedigestedtherecombinantplasmidofpMD18TApxIAandpMD18一TTbpB，thefragmentofApxlAandTbpBgenegot，andthefragmentinsertedtheprokaryotice