RNApolymeraseinprokaB福+Cβaβ'OCoreEnzymeHoloenzymeforelongationforinitiation依靠静电作用力非专一依靠空间结构专一性性与非特异DNA序列结与特异DNA序列结合年衰期；60”半衰期；数小时
RNA polymerase in prok. Core Enzyme • for elongation for initiation • 依靠静电作用力非专一 性与非特异DNA序列结 合 依靠空间结构专一性 与特异DNA序列结合 • 半衰期；60’ 半衰期；数小时 Holoenzyme α β β’ α α ＋ σ α β β’ σ
Core0C(a)tighterbindingHoloenzymeB sitedDElongationcomplexLooselybindingtranscriptionR site
Loosely binding R site tighter binding B site transcription
afactor;★Re-usableafter2-9Nttranscripted★ENSURE:holoenzymerecognizingR sitebindingtemplatestrandofBsite★modify configurationofRNApol降低全酶与DNA的非专一性结合力增强全酶与R，Bsite的专一性结合力导致RNA链的延伸缓慢！
s factor； ★ Re-usable after 2-9 Nt transcripted ★ ENSURE: holoenzyme recognizing R site binding template strand of B site ★ modify configuration of RNApol 降低全酶与DNA的非专一性结合力 增强全酶与R, B site的专一性结合力 导致RNA链的延伸缓慢!
Hybridization-competition(EkkehardBautz)被标记的RNA与相应的非标记RNA（competitor）对同一杂交对象可表现为"competition""competition”3效应越明显表明被标记的RNA特异性越高
Hybridization-competition (Ekkehard Bautz) • 被标记的RNA与相应的非标记RNA（competitor） 对同一杂交对象可表现为“competition” • “competition” 效应越明显 表明被标记的RNA特异性越高
DenatureDNATranscriptionofregionXwith labeled NTPs(＊NTP)aHvrdnonwithno comipetitorHybridizationwith competitorDUnlabeledcompetitorXRNASMeasure labeled hybridsMoasure labelod mybridsRNA competitor★RNAisLabelLabelunlabeledYcompetitortranscribedininnocompetitionhybridhybridfrom region XX RNA competitorAmount oflabeledAmount of unlabeledRNA addedRNA added
unlabeled Y competitor no competition RNA is transcribed from region X Amount of labeled RNA added Amount of unlabeled RNA added