基因表达分析中的内参照设置X

基因表达分析中的内参照设置内参照基因的作用内参照基因的条件成功的实验设计要求的对照实验阳性模板对照阴性对照阳性基因对照内参照阳性基因对照的阴性对照内参照基因的概念和特点所谓内参照,就是实验材料本身固有的,在实验的全程与目的基因相伴随,接受同样的处理,并可作为参照的基因.–特点1:在材料受到不同的(实验处理因素)过程,表达恒定,–特点2:在动物的各种组织中表达比较恒定,–特点3:除实验设计处理因素外,能够与目的基因一起受到同等程度的非设计其他因素影响外参照基因,人工加入的质控成分–不管是何时加入的都是外参好的内参的额外要求表达丰度与目的基因接近在各个物种中序列较为保守,突变机会小功能重要最好是管家基因能够在多种检测中通用操作方便,结果漂亮,重复性好常用内参基因ACTB(actin,beta)GAPDH16Sr18SrTFRC(CD71)TBP(TATA-BOXBINDINGPROTEIN)B2M(BETA2MICROGLOBULIN)PGK1(phosphoglyceratekinase1)GAPDHGlyceraldehyde-3-phosphatedehydrogenase:catalyzesanimportantenergy-yieldingstepincarbohydratemetabolism,thereversibleoxidativephosphorylationofglyceraldehyde-3-phosphateinthepresenceofinorganicphosphateandnicotinamideadeninedinucleotide(NAD).Theenzymeexistsasatetramerofidenticalchains.AGAPDpseudogenehasbeenmappedtoXp21-p11and15GAPD-likelocihavebeenidentified.人GAPDH在染色体上的位置GAPDH在各种组织中的表达水平---电子Northern结果GAPDH在各种组织中的表达水平B2MBeta-2-microglobulinisaserumproteinfoundinassociationwiththemajorhistocompatibilitycomplex(MHC)classIheavychainonthesurfaceofnearlyallnucleatedcells(Gussowetal.,1987[PubMed3312414]).[suppliedbyOMIM]人B2M在染色体上的位置B2M在各种组织中的表达水平---电子Northern结果B2M在各种组织中的表达水平B2M在各种组织中的表达水平-SAGETFRC（CD71）TRANSFERRINRECEPTORFunction:Cellularuptakeofironoccursviareceptor-mediatedendocytosisofligand-occupiedtransferrinreceptorintospecializedendosomes.Endosomalacidificationleadstoironrelease.Theapotransferrin-receptorcomplexisthenrecycledtothecellsurfacewithareturntoneutralpHandtheconcomitantlossofaffinityofapotransferrinforitsreceptor.Transferrinreceptorisnecessaryfordevelopmentoferythrocytesandthenervoussystem(Bysimilarity).Asecondligand,theheditaryhemochromatosisproteinHFE,competesforbindingwithtransferrinforanoverlappingC-terminalbindingsite人TFRC在染色体上的位置TFRC在各种组织中的表达水平TFRC在各种组织中的表达水平---电子Northern结果TFRC在各种组织中的表达水平-SAGEMT-RNR2mitochondriallyencoded16SRNA18SribosomalRNALOC100008588geneThesequencescodingforribosomalRNAsarepresentasrDNArepeatingunitsinthep12regionofchromsomes13,14,15,21and22,designatedRNR1throughRNR5.EachrDNArepeatingunitconsistsofvariablenumbersof5.8S,18Sand28Sgenes.Theseribosomalrepeatingunitsarenotcurrentlyannotatedonthereferencegenome.TBP：TATABOXBINDINGPROTEINInitiationoftranscriptionbyRNApolymeraseIIrequirestheactivitiesofmorethan70polypeptides.TheproteinthatcoordinatestheseactivitiesistranscriptionfactorIID(TFIID),whichbindstothecorepromotertopositionthepolymeraseproperly,servesasthescaffoldforassemblyoftheremainderofthetranscriptioncomplex,andactsasachannelforregulatorysignals.TFIIDiscomposedoftheTATA-bindingprotein(TBP)andagroupofevolutionarilyconservedproteinsknownasTBP-associatedfactorsorTAFs.TAFsmayparticipateinbasaltranscription,serveascoactivators,functioninpromoterrecognitionormodifygeneraltranscriptionfactors(GTFs)tofacilitatecomplexassemblyandtranscriptioninitiation.ThisgeneencodesTBP,theTATA-bindingprotein.AdistinctivefeatureofTBPisalongstringofglutaminesintheN-terminal.ThisregionoftheproteinmodulatestheDNAbindingactivityoftheCterminus,andmodulationofDNAbindingaffectstherateoftranscriptioncomplexformationandinitiationoftranscription.MutationsthatexpandthenumberofCAGrepeatsencodingthispolyglutaminetract,andthusincreasethelengthofthepolyglutaminestring,areassociatedwithspinocerebellarataxia17,aneurodegenerativedisorderclassifiedasapolyglutaminedisease.人TBP在染色体上的位置TBP在各种组织中的表达水平TFRC在各种组织中的表达水平---电子Northern结果TBP在各种组织中的表达水平-SAGE