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. 2018 Mar 2;8:3936. doi: 10.1038/s41598-018-22012-6

Figure 1.

Figure 1

Genomic locus and deletion of Rv3484 from the genome of Mtb H37Rv. (a) Genomic organization of the Rv3484 locus including restriction sites and localization of the probe for Southern analysis and localization of primers for PCR analysis. The in frame deletion comprises 1233 bp. Southern blot of the Mtb H37Rv Rv3484 deletion mutant. (b) Southern analysis using StuI treated genomic DNA resulting in signals at 3878 bp for the mutant strain, two signals at 3878 bp and 4264 bp for the strain which underwent a single crossover resulting in cointegration of a deleted copy of Rv3484 and 5111 bp for the wild type. pSvM2 cut with PacI served as a control and resulted in a fragment of 2266 bp. PCR analysis of the Mtb H37Rv Rv3484 deletion mutant. (c) Agarose gels showing Mtb H37Rv wild type (wt), the strain carrying a deletion in Rv3484 (ΔRv3484) besides the complemented mutant strain (ΔRv3484::Rv3484), the mutant strain carrying the empty vector used for complementation (ΔRv3484::pMV306) and the plasmid used for transformation of the deleted copy of the gene (pSvM2) and a PCR negative control ((−)) as controls. Primers #172 and #185: wt 1481 bp, ΔRv3484::Rv3484 1481 bp. Primers #184 and #190: wt 2913 bp, ΔRv3484, ΔRv3484::Rv3484, ΔRv3484::pMV306, pSvM2: 1680 bp.