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. 2015 Feb 17;43(5):2615–2624. doi: 10.1093/nar/gkv105

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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. — TEFM strongly promotes POLRMT processivity on longer templates. (A) Cartoon of the templates used, produced from pUC18 containing an HSP or LSP insert (mtDNA insert). Note that only the LSP template contains the CSB region. To create run-off products of different lengths, the plasmids were cleaved with different restriction enzymes (RE) as specified in materials and methods. (B) In vitro transcription from LSP (lanes 1–4 and 9–10) or HSP (lanes 5–8) templates. These templates generate transcripts of about 400 nts (lanes 1–2 and 5–6), 3000 nts (lanes 3–4 and 7–8) as shown in panel (A), or 2000 nts (lanes 9–10, LSP PCR fragment template). TEFM was added at 40 nM where indicated. Note that only the LSP templates contain the CSB region, explaining the indicated CSB II-terminated bands present in LSP templates in the absence of TEFM.