FIGURE 4: Role of alr2153 and alr3242 in heme uptake. (A) Strategy model for plasmid insertion in Anabaena genome to generate AFS-I-hutA1 and AFS-I-hutA2. (B) Genomic DNA isolated from Anabaena wild-type, AFS-I-hutA1 and AFS-I-hutA2 was used for PCR with gene-specific primers (lane 1) or plasmid-specific and gene-specific right (lane 2) or left internal primers (lane 3). (C) 10 µl of a culture of Anabaena wild-type (lane 1), AFS-I-hutA1 (lane 2) or AFS-I-hutA2 (lane 3) with a density OD_750nm of 1 or 0.1 as indicated were spotted on solid YBG11 with (upper panel) or without iron (panel 2-4) supplemented with Fe-citrate (panel 3) or Fe-EDTA (panel 4). Representative images of plates after incubation for seven days are shown. (D) The three strains were grown in liquid YBG11 supplemented with 10 mM NaHCO₃ and bubbled with CO₂-enriched air (1% CO₂ v/v). The filament length was counted for 75 filaments in three independent cultures. The bin size is indicated on the right bar. (E) Growth of the three strains in YBG11 (left) or YBG11 –Fe (right) was monitored at OD_750nm after inoculation with a starting OD_750nm of 0.02. The color code is given on the left plot. (F) The three strains were grown for seven days in YBG11 (top) or YBG11 –Fe (bottom) and cultures at OD_{750 nm} = 1 are shown. (G) Growth of the three strains in YBG11 –Fe with 30 µM heme was monitored at OD_750nm after inoculation with a starting OD_750nm of 0.02. The color code is given in (E).