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. 2014 Jun;160(Pt 6):1125–1133. doi: 10.1099/mic.0.077180-0

Fig. 1.

Fig. 1.

Construction of promoter-containing Mtb recombinant strains. (a) Illustration of the Mtb senX3-regX3 2CRS and the strategy utilized to generate promoter-containing recombinant strains. The senX3-regX3 IR consists of two identical 77 bp (white arrows) and a 53 bp (black arrow) MIRUs. (b) The 5′-UTR of senX3 (shaded arrow) was cloned into the promoter-less plasmid pYUB76 upstream of the reporter gene lacZ, generating pSR. A similar cloning strategy was used for the senX3-regX3 IR, yielding constructs pIR-2, pIR-1 and pIR-0.5 containing two full 77 bp MIRUs, one 77 bp MIRU and one 53 bp MIRU, respectively. (c) Nucleotide sequences are shown of the full 77 bp MIRU (white arrow) and the 53 bp MIRU (black arrow) within the senX3-regX3 IR of wild-type CDC1551. Underlined sequences are common sequences shared by MIRUs and bold type indicates putative DTGA insertion sites.