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. 2020 Feb 4;11:684. doi: 10.1038/s41467-020-14508-5

Fig. 3. Deletion and integration of the TbD1 locus in the 79119 and H37Rv genetic backgrounds, respectively.

Fig. 3

a Schematic representation of the genomic organization of the TbD1 locus in the 79112∆TbD1 mutant and complemented strain (79112∆TbD1-C). The schematic representation of the genomic organization of recombinant H37Rv strains, harboring an intact TbD1 locus is also depicted. Arrows indicate primers used in PCR reactions. b, c Amplification profiles obtained in PCR reactions performed on genomic DNAs from different Mtb strains by using primer pairs, specific for TbD1 flanking region. The 2632 bp-fragment obtained in 79112∆TbD1, or the 2632-bp and 3315-bp amplification products detected in 79112∆TbD1-C confirmed the TbD1 deletion and its replacement by a kanamycin resistance gene in the 79112∆TbD1 mutant, the correct TbD1 deletion/re-integration in the 79112∆TbD1-C complemented derivative strain, respectively (b). Similarly, the 1204-bp product obtained in H37Rv, or the 1204-bp and 3330-bp fragments observed in H37Rv::TbD1 clones correspond to the TbD1-deleted locus originally present in WT H37Rv and to the full-size TbD1 region (mmpS6/mmpL6) integrated into the genome, respectively (c).