分子生物学 第4章学习资料 .ppt

b)GroupIsplicingmodel(tetrahymera35srRNAmodel)●intron内含有4个序列保守，两两同源的CCS(centralcoresequence)Junctionsequence.●5’exonUintronGexon3’高度保守，转酯反应的攻击位点形成RNA分子内的二级结构donorreceptorPQRSl?靠近5’junctionseq.的intron内，存在一段与5’,3’两个junctionseq.可发生互补配对，形成二级结构的区域。Internalguidesequence(IGS)SplicingmodelCCS;P/Q,R/SpairingIGS;linktwojunctionseq.trans-esterificationbetweenGU→cutintron,ligationexonsRNAauto-splicingasRibizyme(核酶,核酸质酶)（自身具有酶活性的RNA分子）ExonUGExonIntronS(2)R(9L)PqCCSSplicingmechanismofIntrongroupIIGSc)GroupIIsplicingmodel(Michel1982)JunctionsequenceBranchSiteof7Ntupstreamof3’-junctionseq.●5’exonGUGCGB.SPyAUexon3’供点受点PyPuPyPyUAPy100%●在BranchSite的两侧存在一些短的序列，与其上游互成IR,形成茎环结构(但A不包含在IR序列内,从而被排除成芽状突起）5’--GUGCGPyPuPyPyUAPyPyAU—3’IntronIRIR5’GAU3’ASplicingmechanismIntronIRIRA5’GAU3’Exon1Exon2GAintronPyAUMatureRNA转酯攻击位点intronXOHAU2‘A3‘GCowboy’slasso“Lariat”intronEvidencefortheLariatintermediate1984Sharp4%agar.Lariatintron10%agar.LariatintronE1I1E2TranscriptioninvitroPre-RNA231NtAllowingsometimeforsplicingd)GroupIIIsplicingmodel(Chambon)JunctionsequenceChambonrule●5’exonGUintronAGexon3’供点100%94%受点●7Ntbranchsiteinupstreamof3’junctionseq.pyXpyUpuApy5’E1AAUAGGUGAI1UACAGGUUGE2--E2--CUCAGGUACAI2UCAGGUUGE3--E3CCAGUAAI3UACAGGAAE4--E4AUGGUAA14AAAGGUE5--E5GAGGUAUAUI5CCAGCAAE6--E6GCAGGUAUGGI6GCAGCUUE73’●Intron与Exon的连接处存在部分同源的consensusseq.OvalbuminmRNA(6Intron,7Exon)C