transcript_analyses.ppt

《transcript_analyses.ppt》由会员上传分享，免费在线阅读，更多相关内容在教育资源-天天文库。

1、TranscriptanalysisandreconstructionBrazil2001GenesWhyarethereonlyafewtensofthousandsofgenesinthehumangenome?Howdogenesexpressthemselvestomanufacturetheproteome?Howcanavailablesequenceinformationbeprocessedinordertodeliverunderstandingofgeneexpression?Genomicex

2、pressionWithineukaryotes,geneshavesharedbasiccharacteristics.Theyhavesingleormultipleexonsandintronsdistributedalongthegeneincodingandnon-codingregionswith5’FlankingregionwithtranscriptionregulationsignalsTranscriptioninitiationstartsite(5’)Initiationcodonforp

3、roteincodingsequenceExon-intronboundarieswithsplicesitesignalsattheboundariesTerminationcodonforproteincodingsequence3’signalsforregulationandpolyadenylationGCboxCAATTATAGCboxTranscriptionInitiationSiteExon1InitiationCodonGTTranscriptionInitiationSiteAGGTIntro

4、n1Intron2Exon2GCboxGCboxCAATTATATranscriptionInitiationExon1InitiationCodonGTAGGTAGIntron1Intron2Exon2Exon3StopCodonAATAAPoly(A)additionsite5’FlankingregionPre-mRNAMaturemRNAGeneExpressionTranscriptionproductscanvary.Transcriptioninitiationatthestartsite(TSS)E

5、xonlengthExonprescence/absenceinthematuretranscriptAlternatetranscriptionterminationandpolyadenylationAlternativedonorandacceptorsplicesitesAlternativepolyadenylationExonskippingExamplesofalternativesplicingGCboxGCboxCAATTATATranscriptionInitiationExon1Initiat

6、ionCodonGTAGIntron1Exon3StopCodonAATAAPoly(A)additionsite(s)Exon2SKIP3’FlankingregionPre-mRNAMaturemRNACapturingexpressedtranscriptsDatabases-SequencesdbESTSeveralcollapseddatasetsTIGR-THCAllgenesUnigeneBodyMapSTACKSeveralmorespecialisedGenomeSequenceasitappea

7、rs:ExpressionCaptureSerialAnalysisofGeneExpressionDNAfragmentsthatactasuniquemarkersofgenetranscripts.AssayofnumbersofeachmarkerinasetofsequenceyieldsameasureofgeneexpressionArrayLaydownofsequenceclonestoprovideanorganisedseriesforhybridisationResolutionofCapt

8、uredExpressionESTSLowresolution,broadcapture,providestemplateforSAGEandArraySAGEMediumresolution,needtemplate,noisecanbeanissue,stoichiometryisrevealedbutstandardisationaproblemARR