16403.非洲爪蟾april及犬light、hvem的克隆、表達及生物學活性研究

1、學位論文獨創(chuàng)性聲明本人鄭重聲明：所提交的學位論文是本人在導師指導下進行的研究工作和取得的研究成果。本論文中除引文外，所有實驗、數據和有關材料均是真實的。本論文中除引文和致謝的內容外，不包含其他人或其它機構己經發(fā)表或撰寫過的研究成果。其他同志對本研究所做的貢獻均已在論文中作了聲明并表示了謝意。學位論文作者簽名：舅j倭日期：2啾、s、≯歲學位論文使用授權聲明研究生在校攻讀學位期問論文工作的知識產權單位屬南京師范大學。學校有權保存本學位論文的

2、電子和紙質文檔，可以借閱或上網公布本學位論文的部分或全部內容，可以采用影印、復印等手段保存、匯編本學位論文。學校可以向國家有關機關或機構送交論文的電子和紙質文檔，允許論文被查閱和借閱。(保密論文在解密后遵守此規(guī)定)保密論文注釋：本學位論文屬于保密論文，密級：——保密期限為——年。學位論文作者簽名：努)霞日期：Mk畎指導教師簽名：限。a／令日期：】挑、g諤AbstractAbstract’IhethesiscOntainstwOchapt

3、crs：SectiononeInthisstudy，aX認P融Lh鵲beenclonedaIldidentified舶mA仔icanclawed舶gbyR●PCRaIldRACEtecIlIlologyThemlllengthcDNAofXlAPⅪLwas1504bp，consistingofa91bpby5’一tenninaluntanslatedregion(UTR)，a714bpby3’te唧inaluntallslatedreg

4、ionaIlda699bpopenreading仔鋤e(ORF)encoding264aminoacidsXL心剛Lcon協inst、Ⅳocysteineresidues(Cysl78aIldCysl93)，a如rinpmte雒ecl∞VagesiteandatypicalTNFhomologydomaincorrespondingt0otheridentifiedAPRILhomologuesSequencealignmentshow

5、sthatthesequencehomologyishigherintheex仃牝ellulardomainthanintheintraceIlulardomain，whichindicacesthatthereceptorbirldiIlgdomainismoreconservedthanthec)喲plasmic協ilRea㈨mequ鋤titatiVeRTPCRdemonstratedhigheXpressionofXlAPRILj

6、nthemairIimmuneo唱anspleenSUMoIXsAPRIL如sionproteinwasemcientlyexpressedinEcD療Rosseta(DE3)andpurifiedwithNi—N1’Aaffinitychmmatogmphy111ent11eexpressionofsuMOxsAPⅪLwasanaIysedbySDSPAGEandrecogllizedbyWestemblottingusing蛐鋤ti

7、HiS6tagmouseantibodyRecombinantxsAPRjLcouldpromotethesurviva師roIiferationof仔oglymphocytesaIldmousesplenicBcells加vf拋SectiontwoLIGHT鋤dHVEMareimportantmembersoftheTNFfamilyaIldTNFRFf鋤ily，respectiVelyDuetotheimportancemlesin

8、illlmunoregulation，theLI(HTgeneof723bpandHVEMof819bpwere鋤plifiedfh)mdogspleenbyImPCRinthepresentstIldyBioinfomatics柚alyseswereconductedtodescribegeIlcsstmctIlrecharacteristicswhichsuggestedthatLIGHTwasat)，peII仃ansm鋤brane

9、proteinandHVEMwasa鑼peItransmembraneproteinTheresultsofreal—timePCRreVealedthatbothdLIGHTanddHVEMwerceXpressedinvarioustissuesandelevatedinimmuneorgans‘IherclativeexpressionlevelsofdLIGHTaIlddHVEMwerethehighestinspleenWbh

10、aVecons打uctcdtwecombinantplasmidswhichn鋤edaspET431枷sLIGHTandpET28a—dsHVEMtIlrou曲geneticengineeringmethodsFusionproteinwasemcientlyexpressedinEco，fBL2l(DE3)andthenpurifiedwithNi—N‘【Aaffinit)rchromatographySomepilotstudies