柔嫩艾美耳球蟲楊凌株cDNA文庫(kù)的免疫學(xué)篩選及其3-1E基因的克隆與表達(dá).pdf

1、、，西北農(nóng)林平杖大學(xué)2007屆攻讀碩士學(xué)位研究生學(xué)位(畢業(yè)j論文柔嫩艾美耳球蟲楊凌株cDNA文庫(kù)的免疫學(xué)篩選及其3—1E基因的克隆與表達(dá)學(xué)科、專業(yè)研究方向研究生指導(dǎo)教師合作指導(dǎo)教師完成時(shí)舊：塑陵置醫(yī)生一j絲赴勤韭蝴iL一摧芏星i：圣杜熬握』芏螋盟紅曼2007印3月蘭一裝ⅫIMMUNoSCREENINGOFcDNAEXPRESSIONLIBRARYOFE勱刃曼足丟4乃酣憶￡厶4YLSTRAINANDCLONINGEXPRESSIONOFM

2、AJORANTIGENGENEABSTACTChickencoecidiosisisallimportantaviandiseasewhichseriouslyharmsthepoutryindustryItwascausedbyvariousEimeriasppparasitizedinintestinalmucosalepitheliawhicharecharacterizedwithentericpathologicallesio

3、nsAmongvariousEimeriaspp。，Etenellawhichca髑escaccalcoccidiosisishighlypathogenicPresengtlythecontrolofcoccidiosischieflydependsuponprophylacticchemotherapywimanticoeeidialdrugsHowevertheemergenceofdrugresistanceincoceidia

4、isagreatproblemwitIImostofthedrugs，whichinduecour跎，limitstheiruseFurthermore，drugorantibioticresidueinthepoultryproductispotentiallyharmfultoconsumersTheselimitationshavenecessitatedthesearchforalternativeEimeriacontrolm

5、easuresAmongtheseveralalternativeEimeriacontrolmeasllrasonlyhostvaccinationagainstEimeriaappearspromisingInthisrcsearch，ourstrategywasscreeningtheconstructedcDNAexpressionlibraryofthesporozoitesbyEtenellaYLstrainsporulat

6、edoocystsbyimmunologicalmethods，cloningandexpressingtheantigengenes，analyzingtheprotectiveimmunityResultsofstudiesdisplayedasfollowing1eDNAexpressionlibraryoftheEtenellaYLstrainsporozoiteswasimmunoscrcenedwithpositiveser

7、aofchickenanti—EtenellaandmbbRanti—EtenellaYLstrainThus3positivecloneswereobtained，onepositiveclonegenewasdesignated，andtheORl：ofthisgeneisconsistedof513nucleotidesencoding170aminoacidsTherewerellighhomologieswithpublish

8、ed31EgeneingenBankComparedwith3IEofEtenellaGSstrainandUS蚰咖thehomologyofnucleotidesacidsequenceofEtenellaYLstrain3IEwere998％and994％respectivelythehomologyofaminoacidencodedwere988％and98_2％，respectivelyAllthoseconfirmedtha

9、tthegenewasanewgeneofEtene／／anenewgeneaccessionnumberinGenBankisEF069437anditwasnamedasEimeriatenellastrainYL31E2Theinterestinggeneof31EfloralibrarywasdonedtoexpressionvectorofpGEX4T=1withspecialprimer11∞fusionexpression

10、vectorofpGEX03一lEandwasconstructedthentransformedintoEcoliBL21Thefusionexpressionof31EwasinducedwithIPTGtheexpressionproductwasanalyzedbySDSPAGEmresultsshowedthatthe31EfusionproteinwasexpressedsuccessfullyinBL21mfusionpr