TheMatchmakerGoldYeastTwo-HybridSystemisthemostadvancedandhighestperformingversionofourhighlypopularMatchmakerproductlineforinvestigatingprotein-proteininteractions.Thisisduetoacombinationofanewyeaststrain(Y2HGold),stringentreporters,easy-to-uselibraries,high-levelexpressionvectors,andSMART-basedlibraryconstructionkits.
TheMatchmakerGoldYeastTwo-HybridSystemisthemostadvancedandhighestperformingversionofourhighlypopularMatchmakerproductlineforinvestigatingprotein-proteininteractions.Thisisduetoacombinationofanewyeaststrain(Y2HGold),stringentreporters,easy-to-uselibraries,high-levelexpressionvectors,andSMART-basedlibraryconstructionkits.
OverviewofMatchmakeryeasttwo-hybridassay
InaMatchmakerGAL4-basedtwo-hybridassay,abaitproteinisexpressedasafusiontotheGAL4DNA-bindingdomain(DNA-BD),whilelibrariesofpreyproteinsareexpressedasfusionstotheGAL4activationdomain(AD)(Chienetal.1991;FieldsandSong1989).Whenbaitandlibrary(prey)fusionproteinsinteract,theDNA-BDandADarebroughtintoproximitytoactivatetranscriptionoffourindependentreportergenes(AUR1-C,ADE2,HIS3,andMEL1).
Thistechnologycanbeusedto:
- Identifynovelproteininteractions
- ConfirmsUSPectedinteractions
- Defineinteractingdomains
WhydoIneedfourreportergenes?
Yeasttwo-hybridsystemsthatutilizeonlytheHIS3nutritionalreportertoscreenforprotein-proteininteractionsoftengenerateahighnumberofbackgroundcoloniesandfalsepositives.Backgroundcoloniesaretheresultofleakyexpressionofthenutritionalreporter,whereasfalsepositivesarecausedbypreyproteinsthat,independentlyofthebait,havetheABIlitytorecognizeandbindtosequencesupstreamofthereporter.
TheMatchmakerGoldYeastTwo-HybridSystemistheonlyyeasttwo-hybridsystemwithfourgenuinereportersandthreedifferentbaitrecognitionsequences.BackgroundgrowthisnotpossIBLeduetothepotencyandstabilityofAureobasidinA,anovelyeastantibioticthatkillsallnon-resistantcells.Falsepositivesarereducedtoaminimumbecauseanyindependentlybindingpreyproteinwouldneedtobindtoallthreerecognitionsequencesandactivatefourdifferentreporters.
Thefourreporters
Thesystemutilizesantibiotic,nutritionalandcolorimetricreporters.
Yeasttwo-hybridmediasets
Ouryeasttwo-hybridmediasetseachcontainacompletesetofthealltheyeastmediapouchesyouneedforMatchmakerGoldtwo-hybridprotocols.
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IgM是免疫球蛋白M(Immunoglobulin M，IgM)的缩写。根据结构的不同将免疫球蛋白分为五种，IgM是人的免疫球蛋白之一，其他还有lgA、lgG、IgD和lgE。
HSV1型导致的一般都是腰部以上的感染，典型症状是水泡有可能引起脑炎，而且有非活化状态的潜伏。所以我觉得你根本就不用担心这个抗体IgG阳性的化验单。
2-IgM也是一种特异性感染指标，但在人体内持续时间不长（有时是30天更短），往往是传染病的近期感染指标。呈阴性是指：你近期没有感染过结核杆菌。
文章原文链接http://www.jbc.org/content/282/23/16776.long
