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. 2015 Oct 20;43(21):10411–10420. doi: 10.1093/nar/gkv1095

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — In vitro transcription by phiKZ nvRNAP. (A) Multi-round run-off in vitro transcription by nvRNAP from PCR fragments containing an early (P_54E), middle (P_62M) and late (P_119L) phiKZ promoters. RO – run-off RNA products. (B) Primer extension mapping of 5′-ends of in vivo (left panel) and in vitro (right panel) transcripts from the phiKZ late promoter P_119L. DNA sequencing reactions with the same end-labeled primer used as sequence markers are indicated. The arrows indicate primer extension products. (C) In vitro abortive initiation reactions by nvRNAP and host P. aeruginosa RNAP σ⁷⁰-holoenzyme (Pa RNAP) from phiKZ late promoter (P_119L) and bacterial RNAP promoter T7 A1. 3 nt – trinucleotide abortive transcripts. (D) In vitro run-off transcription by nvRNAP and P. aeruginosa RNAP from phiKZ P_119L and T7 A1 promoters in the presence or absence of rifampicin. RO – run-off RNA products.