Figure 5

Complementation of csaB mutation. a) PCR verification of XmaI mutant csaB. The wild-type sequence of csaB was restored in the ΔcsaB strain with the addition of a silent mutation creating an XmaI site (see Methods). The region was amplified by PCR, and PCR products were purified and digested with XmaI. The lane labeled BA663 indicates the csaB PCR product from wild type strain (7702) after digestion with XmaI, whereas the lane labeled BAP533 is the PCR product from the mutant showing the two bands after digestion with XmaI. b) Genetic map of csaB complementing plasmid pNBGD1003. pNBGD1003 is a shuttle vector derived from pSW4. The locations of the Gram positive and Gram negative replicons (from pUB110 and ColE1, respectively), antibiotic resistance genes aadD and bla, origin of conjugational transfer oriT, constitutively expressed B. anthracis pagA promoter (labeled PA promoter) and the csaB gene downstream of the promoter are indicated.