16403.非洲爪蟾april及犬light、hvem的克隆、表達(dá)及生物學(xué)活性研究

1、學(xué)位論文獨(dú)創(chuàng)性聲明本人鄭重聲明：所提交的學(xué)位論文是本人在導(dǎo)師指導(dǎo)下進(jìn)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(shù)據(jù)和有關(guān)材料均是真實的。本論文中除引文和致謝的內(nèi)容外，不包含其他人或其它機(jī)構(gòu)己經(jīng)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(xiàn)均已在論文中作了聲明并表示了謝意。學(xué)位論文作者簽名：舅j倭日期：2啾、s、≯歲學(xué)位論文使用授權(quán)聲明研究生在校攻讀學(xué)位期問論文工作的知識產(chǎn)權(quán)單位屬南京師范大學(xué)。學(xué)校有權(quán)保存本學(xué)位論文的

2、電子和紙質(zhì)文檔，可以借閱或上網(wǎng)公布本學(xué)位論文的部分或全部內(nèi)容，可以采用影印、復(fù)印等手段保存、匯編本學(xué)位論文。學(xué)?？梢韵驀矣嘘P(guān)機(jī)關(guān)或機(jī)構(gòu)送交論文的電子和紙質(zhì)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(xué)位論文屬于保密論文，密級：——保密期限為——年。學(xué)位論文作者簽名：努)霞日期：Mk畎指導(dǎo)教師簽名：限。a／令日期：】挑、g諤AbstractAbstract’IhethesiscOntainstwOchapt

3、crs：SectiononeInthisstudy，aX認(rèn)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(xié)inst、Ⅳocysteineresidues(Cysl78aIldCysl93)，a如rinpmte雒ecl∞VagesiteandatypicalTNFhomologydomaincorrespondingt0otheridentifiedAPRILhomologuesSequencealignmentshow

5、sthatthesequencehomologyishigherintheex仃牝ellulardomainthanintheintraceIlulardomain，whichindicacesthatthereceptorbirldiIlgdomainismoreconservedthanthec)喲plasmic協(xié)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