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. 2017 Aug 22;12(8):e0183346. doi: 10.1371/journal.pone.0183346

Fig 3. Creation of B. anthracis strains containing variable numbers of atxA genes replacing the nprC gene.

Fig 3

(A) Plasmids used for sequential atxA gene insertions. (B) Scheme indicating sequential atxA gene insertions: nprC gene was replaced by the first atxA gene using Flp-FRT system with pSCF-2183LA plasmid as a donor of atxA; the second atxA was added from pSC-A plasmid with Cre-loxP system; and the third atxA copy was inserted with IntXO-PSL system from pSCP-A plasmid. (C) PCR confirmation of atxA gene insertions into genome of B. anthracis (primers 2183seqF/2183seqR). Strains with one, two, and three copies of atxA are indicated on the top of the gel. (D) PCR confirmation of atxA intergenic regions in strains with double and triple atxA genes (primers seqALAF/ALARseqR).