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. 2013 Dec 15;3(1):29–41. doi: 10.1002/mbo3.145

Figure 2.

Figure 2

The strategy used to disrupt parB in Xac. The suicide vector pPAU2, which carries the parB124–769 fragment, was integrated into the parB locus of Xac by a single crossover event; integration generated the genomic structure depicted underneath the vector in which ParBΔ52 can be expressed under the control of the native promoter of ParB (pparAB). (A) Genomic coordinates of parAB. (B) Schematics of integration: upon plasmid insertion, two new forms of parB will be generated: one coding for ParBΔN41, a fusion between the first 26 amino acids of LacZα (coded by the expression vector) plus residues 42–308 of ParB, and another that codes for ParBΔC52, which lacks the 52 C-terminal residues of ParB and has an addition of the 18 new amino acids shown. Both truncated ParB proteins still carry the helix-turn-helix (HTH) motif shown. The numbers above the map and around the circle indicate the sizes of the DNA fragments delimited; the predicted helix-turn-helix motif of ParB is depicted below the drawings; ERV, EcoRV restriction sites. (C) Southern blot analysis: total DNA of the three Xac parB::pAPU2 mutants was digested with EcoRV, and probed with parB. The sizes of the hybridization fragments detected correspond to those estimated based on the genomic coordinates shown in B.