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. 2010 Jul 16;192(18):4562–4570. doi: 10.1128/JB.00536-10

FIG. 3.

FIG. 3.

The two copies of the 350-kb duplicated region are arranged in tandem. (A) A schematic representation of the duplicated region is shown. Genes Rv3128c and Rv3427c mark the approximate beginning and end of each duplicated segment, respectively. Genes Rv3127c and gadB flank the duplication externally. The length of the duplication is indicated in parentheses, and the two copies (Copy 1 and Copy 2) are arranged in a direct, tandem duplication. The arrangement of the genes indicated (boxed) was confirmed by Southern blotting of genomic DNA comparing wild-type H37Rv and the W/Beijing isolates HN878 and G4B1.2. The approximate locations of the restriction enzyme sites are based upon the published H37Rv sequence. (B and C) Southern blots confirming that the beginning of the duplication is located between Rv3127c (single copy) and tgs1 (duplicated). DNAs were digested with XbaI and hybridized with a 620-bp probe derived from Rv3127c (B; primers Rv3127c-A and -B) or with a 920-bp tgs1 probe (C; primers Rv3130-1 and Rv3130c-F). (D and E) Southern blots confirming the location of the junction of the duplication and that the two copies are arranged in a direct, tandem duplication. DNAs digested with HindIII and SspI were hybridized with a 540-bp alr probe (D; primers alr-C and alr-R) or with a 1-kb dosR probe [E; primers dosR-C and dosRrev(HindIII)]. (F and G) Southern blots confirming that the end of the duplication is located between Rv3427c (duplicated) and gadB (single copy). DNAs were digested with BamHI and NheI and hybridized with the 540-bp alr probe (F) or a 1.2-kb gadB probe (G; primers gadB-C and gadB-D).