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. 2017 Oct 25;7:13987. doi: 10.1038/s41598-017-14410-z

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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PafB and PafC form a heterodimeric PafBC complex co-expressed from a single pafABC mRNA. (A) The pafABC operon. Co-transcription of pafABC in Msm was confirmed by isolating mRNA from the wild type strain and a subsequent reverse transcriptase reaction. cDNA was analysed by PCR using oligonucleotides that resulted in the amplification of products spanning either pafA and pafB (AB) or pafB and pafC (BC; as indicated by black bars). PCR reactions using Msm genomic DNA (gD) and mRNA that was not subjected to reverse transcription (−) served as positive and negative control, respectively. (B) Analytical gel filtration of PafB and PafC separately (upper panel) and co-expressed and co-purified PafBC complex (lower panel). For each trace, 30 µM protein was incubated 20 min at 37 °C before running on a Superdex 200 10/300GL column at 1 ml/min at room temperature. Peak fractions analyzed by SDS-PAGE are shown as gel slices above their respective peaks.