FIG 2.
Analysis of cell morphology, growth curves, and plasmid profile of S. aureus RN4220 transformed with shuttle vector pBUS1, pBUS1-Pcap, pBUS1-Pcap-HC, pTSSC-Pcap, and pTSSCm-Pcap. (A) Colony size of RN4220 transformants after 16 h of growth on LB agar containing 10 μg/ml tetracycline. (B) Growth of RN4220 transformants in LB broth containing 10 μg/ml or 2 μg/ml tetracycline monitored through OD600 measurements during 16 h. Strain RN4220 without plasmid inoculated in selective (LB-tet RN4220) and nonselective (LB-RN4220) medium was used as a control. OD600 values are averages from triplicates. Note, OD600 values of 0.2 to 0.3 correspond to dense stationary-phase cultures; low values are due to measurement conditions, including short light path length. (C) Plasmid yield of the different vectors in RN4220. Plasmid DNA was isolated from similar 14-ml overnight cultures grown in selective LB broth and separated on 1% agarose gel. One-tenth of total plasmid extraction was loaded. M, 1 kb DNA ladder (Solis BioDyne).