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1、yolkantibodygroupwassignificantlylowerthanthecontrolgroup.AtthesametimeofenkaryoticexpressionCOEgenecloningtechnologytopichiaexpressionvectorpPIC9K,expressedinpichia,COEforeukaryoticexpressionrecombinantprotein.Westernblottoidentifythreetypesofrecombinantpro
2、teinshaveantigenicity.COEexpressionofrecombinantproteinsineukaryoticemulsifiedwithFreundfsadjuvantimmunizedmiceaftertotalfreetwiceatintervalsoftwoweeks,thefirstfivedoseof60
3、ig/only,twofreedosesof80
4、ig/only.Tendaysafterthesecondimmuniz-ationandbloodtestscarri
5、edoutinScrumneutralizationtiters.TheresultsshowedthatsuccessfullyclonedPEDVS534(534bp)andCOE(530bp)gene,PEDVS534proteinandproteinsizeCOEprokaryoticexpressionwere37.5kDaand37.0kDa,consistentwiththetheoreticalvalue,theexpressionoftheproteinandimmunogenic.Eukar
6、yoticproteinexpressionCOEsizeof27kDa,largerthanthetheoreticalvalue,theproteinmaybeduetotheglycosylationreactionoccurs,theneutralizingantibodytitersinmiceimmunizedwiththeproteinproducedbyexpressionreaches1:36.PEDVS534prokaryoticexpressionofneutralizationtiter
7、PEDVCOEproteinsandwholevirusasanimmunogenpreparedyolkantibodieswere1:12,1:25and1:120.Therecombinantproteinyolkantibodyanti-COEclinicaltrialsshoweddiarrheayolkantibodygroupwas5.56%,whilethemortalityratereached61.11%inthecontrolgroup,Thepigletsdiarrhearateofex
8、perimentalgroupandcontrolgroupwasnotsignificant.InthisstudyPichiaexpressionsystemofPEDVCOE,proteinexpressionandcaninduceneutralizingantibodiesinmice;successfullypreparedthreeyolkantibodies,andthreeeggyolkantibodyshowedsomeabilitytoneutralizethevirus;Theresis
9、tancetoCOErecombinantproteinyolkantibodyhassignificanteffectoncontrolofdiarrheapigletsmortality,forfurtherstudyanduseitasafeedadditivetoprovideatheoreticalbasis.KeyWords:porcinediarrhea;eggyolkantibody;feedadditive;COEgene;neutralizationepitope;eukaryoticexp
10、ressionin目录摘要IAbstractII文献综述11卵黄抗体概述11」IgY的结构11.2卵黄抗体的性质及特点11.2.1产量高11.2.3性能稳定11.2.4治疗效果好，安全性高21.3卵黄抗体的应用21.3.1卵黃抗体在诊断方面的应用21.3.2卵黄抗体在防治疾病方面的应用22卵黄抗体饲料添加剂的研究进展33猪流行性腹泻概述43•1丿-X**扌3：彳iE••••••••••
