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Leading strand replication of pMV158 derivatives initiates at the same site in S. pneumoniae that is cleaved by RepB protein in vitro. Bacteria were cultured to OD600 0.25–0.3. Aliquots containing 4 × 106 cfu of S. pneumoniae strains MP551[pLS86] (lane 1), MP551[pLS1cop7] (lane 2), and MP551[pCGA12] (lane 3) were subjected to runoff DNA synthesis, as described, except that 10 ng of primer pMV158 was present in each reaction mixture.
