FIG 4.

Search for regions involved in stability of pMS10. (A) Construction of the pMS10 deletions. Two stem-loops of 67 and 45 bp were found within pMS10 NaeI/Bsu36I fragment. Deletion of the 67-bp stem-loop resulted in pMS11, while deletion of the 45-bp stem-loop resulted in pMS12. Deletion of the pMS10 Bsu36I/NaeI fragment of pMS12 resulted in pMS13. Two more stem-loops of loops 41 and 55 bp are shown on the plasmid map. These are potential transcriptional terminators for GBAA_pXO1_0069 and GBAA_pXO1_0080, respectively (ORF 80 was deleted during pMR8186 creation). (B) Percentage of spectinomycin-resistant (i.e., plasmid-containing) bacteria in cultures of B. anthracis Ames 33 containing either pMS111, pMS122, or pMS13 were determined as described previously.