Recent publications have shown that active RNA polymerase (RNAP) from Mycobacterium tuberculosis (MtbRNAP) can be produced by expressing all four subunits in a single recombinant Escherichia coli strain [1–3]. By reducing the number of plasmids and changing the codon usage of the Mtb genes in the co-expression system published by Banerjee et al. [1], we present a simplified, detailed and reproducible protocol for the purification of recombinant MtbRNAP containing the ω subunit. Moreover, we describe the formation of ternary elongation complexes (TECs) with a short fluorescence-labeled RNA primer and DNA oligonucleotides, suitable for transcription elongation studies.

This work was supported by Fondecyt 11110534 (to M.B.), 1151274 (to M.B.), 11130263 (to C.A.M.W.), Anillo 1107 (to M.B.), and Chile and Fondecyt 196–2013 (to D.G.G.) CONCYTEC, Perú. This research was partially supported by the supercomputing infrastructure of the NLHPC (ECM-02) at Universidad de Chile. A.B. was supported by Conicyt master fellowship 22121199. We gratefully acknowledge Professor Alexander Vologodskii from New York University for helpful comments and advice. Travel for C.B. to and from Chile and Peru was also partially supported by the Howard Hughes Medical Institute, NIH grant R01GM032543, and the U.S. Department of Energy Office of Basic Energy Sciences Nanomachine Program under contract no. DE-AC02-05CH11231.

Background Evidence regarding differences in survival associated with the site of metastasis in triplenegative breast cancer (TNBC) remains limited. Our aim was to analyze the overall survival (OS), distant relapse free survival (DRFS), and survival since the diagnosis of the relapse (MS), according to the side of metastasis. Methods This was a retrospective study of TNBC patients with distant metastases at the Instituto Nacional de Enfermedades Neoplasicas (Lima, Peru) from 2000 to 2014. Prognostic factors were determined by multivariate Cox regression analysis. Results In total, 309 patients were included. Regarding the type of metastasis, visceral metastasis accounted for 41% and the lung was the most frequent first site of metastasis (33.3%). With a median follow-up of 10.2 years, the 5-year DRFS and OS were 10% and 26%, respectively. N staging (N2-N3 vs. N0, HR = 1.49, 95%CI: 1.04-2...