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. 2023 Dec 18;30:95. doi: 10.1186/s12929-023-00988-2

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© The Author(s) 2023

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 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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Fig. 1 — Generation of the Mycobacterium bovis BCG tbcm deletion mutant. A Schematic diagram of the tbcm gene deletion strategy. B Whole-gel image of polymerase chain reaction (PCR) confirmation of tbcm gene (600 bp) expression in wild-type BCG (BCG) and BΔtbcm. C Whole-genome sequencing result of gDNA isolated from BΔtbcm. The tbcm gene deletion was confirmed by a depth level of 0 in the sequencing depth plot. D Uncropped blot image of TBCM (left) and hsp65 (right) protein expression in BCG and BΔtbcm. rTBCM represents recombinant TBCM. E Graphs comparing the growth curves of BCG (gray) and BΔtbcm (blue). The growth curve of each mycobacterial strain was drawn by measuring the OD₆₀₀ at each time point. All data were collected in at least three independent experiments and are presented as the mean ± SEM. The results were analyzed for statistical significance by two-way ANOVA with Tukey’s multiple comparison test. (*P < 0.05, **P < 0.01, ***P < 0.001). BCG wild-type BCG, BΔtbcm, BCG Δtbcm mutant