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Bacterial strains and growth conditions

AL Andrés León-Torres
EA Epifania Arango
EC Eliana Castillo
CS Carlos Y. Soto
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M. tuberculosis H37Ra was grown in Middlebrook 7H9 or 7H11 supplemented with 10% OADC (50 μg/mL oleic acid, 0.5% Bovine albumin Fraction V, 0.2% dextrose, and 0.004% catalase), or Sauton medium supplemented with 0.2% glucose, 30 μM Fe2(SO4)3, and 0.05% Tween-80. M. smegmatis mc2155 cells were grown in LB or Sauton medium supplemented as described above. Mycobacteria were grown until an OD595nm = 0.4 for electroporation experiments or OD600nm = 0.6 for plasma membrane extraction. Escherichia coli TOP10 and HB101 host cells were cultured in LB and used for plasmid constructions. When needed, 7H9 or 7H11 were supplemented with 10% OADC, 0.05% Tween 80, 20 μg/mL kanamycin (Km), and 50 μg/mL hygromycin (Hyg). Likewise, LB was supplemented with 50 μg/mL Km, 100 μg/mL Hyg, 100 μg/mL ampicillin (Amp), 0.5 mM isopropyl-β-d-thiogalactopyranoside (IPTG), and 80 μg/mL 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside (X-Gal).

Copyright and License information: The Author(s) ©2020
Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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