鴨疫里默氏菌ragB基因的生物信息學(xué)分析和克隆表達(dá).pdf

1、論文獨(dú)創(chuàng)性聲明本人鄭重聲明：所呈交的學(xué)位論文是我個人在導(dǎo)師指導(dǎo)下進(jìn)行研究工作所取得的成果。盡我所知，除了文中特別加以標(biāo)注和致謝的地方外，學(xué)位論文中不包含其他個人或集體已經(jīng)發(fā)表或撰寫過的研究成果，也不包含為獲得四川農(nóng)業(yè)大學(xué)或其它教育機(jī)構(gòu)的學(xué)位或證書所使用過的材料。與我一同工作的同志對本研究所做的任何貢獻(xiàn)均已在論文中作了明確的說明并表示了謝意。研究生簽名：沙Is年6旯l；日關(guān)于論文使用授權(quán)的聲明本人完全了解四川農(nóng)業(yè)大學(xué)有關(guān)保留、使用學(xué)位論文

2、的規(guī)定，即：學(xué)校有權(quán)保留并向國家有關(guān)部門或機(jī)構(gòu)送交論文的復(fù)印件和電子版，允許論文被查閱和借閱，可以采用影印、縮印或掃描等復(fù)制手段保存、匯編學(xué)位論文。同意四川農(nóng)業(yè)大學(xué)可以用不同方式在不同媒體上發(fā)表、傳播學(xué)位論文的全部或部分內(nèi)容?！瘫菊撐牟槐Ｃ?。(請在以上口內(nèi)劃“√”)研究生簽名：拇導(dǎo)師簽名：證謊菊口本論文保密，在一年解密后適用本授權(quán)。blS年笠毛竅|∑日月日四川農(nóng)業(yè)大學(xué)碩士學(xué)位論文AbstractBioinlormaticsAnalysl

3、sanaExoresslon0IKtemereila1n●l●11廣’●一一■■■antipestiferragBanragBgene2YangSisi(PreventiveVeterinaryMedicine)DirectedbyProfessor：WANGMingshuRiemerellaanatipestifer(RA)cancauseducks，geese，turkeysandotherpoultryduckplaguecrea

4、merfungusinfectionThediseaseisakindofisacuteorchronicsepticaftercontactwithsexuallytransmitteddiseases，andiscurrentlyoneoftheimportantdiseasesendangeringtheduckindustryInthisstudy，whatIdidistheragBgene(GenbankaccessionNo

5、：CP00378711fromRACH一1strainfocusesresearchesonbioinformaticsanalysiscloingandexpressionofRAragBgeneThemainresultsareasfollows：1BioinformaticanalysisofRiemerellaantipestiferragBgeneThewholeragBgenelengthisl593bpofRiemerel

6、laanatipestifertheGCcontentofthegenesequencesis3572％ThegenecontainsaopenreadingflameisORFl593Theeffectivenumberofcodons(ENC)is38491111isORFcontains43rarecodons，3codontwocoupletbutnoncontinuousmorethantworarecodonsstringF

7、iftyninecodons(excludingMet，Trpandtheterminationcodons)inthepolypeptide，witheightsynonymouscodonsstrongbiastowardAendedandtwelvetowardTendedatthethirdcodonposition，wereusedThereare25codonsshowingdistinctusagedifferencesb

8、etweenRAragBandEcoli，30betweenRAragBandHomosapiens，26codonsbetweenRAragBandyeastThereforetheyeastandEcoliexpressionsystemmaybesuitablefortheexpressionofRAragBgeneifsomecodonscouldbeoptimizedRARagBencoding530aminoacidprot

9、einwiththeoreticalmolecularweightof5897kDaandisoelectricpointpI5716，whichcontained2Nglycosylationsites，5ProteinkinaseCphosphorylationsites，3CaseinkinaseIIphosphorylationsites，9Ncardamomacylationsite，2Tyrosinekinasephosph

10、orylationsite1cAMPandcGMPdependencesite，and1signalpeptidebutwithouttransmembranedomainAnalysisoftheantigenicproteinindicatesthatthe6077，186201，251269，387～403，495～513aminoacidshasacontinuousstrongantigenicityThesubcellula

11、rlocalizationanalysisshowedthattheproteinismainlydistributedintheoutermembranePhylogeneticanalysisshowedthataminoacidsoftheproteinwithothermembersofFlavobacteriaceaeRagBproteinhavealowsimilarity2ProkaryoticexpressionRAra

12、gBgeneWeusedOligo71designedapairofprimersandusedtoamplifytheragBgenewhichfragmentis1623bplengthApMD19一TragBcloningvectorandapET32aRagBexpressionvectorwereconstructedandexpressedinEcoliBL21systemTheexpressionconditionsis：