FIGURE 3.

anaTic22 is involved in OM biogenesis. A, the genomic structure of anaTic22, including downstream encoded genes, the annealing position of the oligonucleotides, the position of GFP insertion to generate strain AFS-PDGF-tic22 and the position of the plasmid insertion to generate strain AFS-I-tic22 are indicated. B, shown is the PCR on genomic DNA isolated from AFS-I-tic22 (AFS-I) and Anabaena sp. PCC 7120 (WT) using the indicated primer combinations (lanes 1–4). The amplification of the rnpB region was used as loading control. C, immunodecoration of cell lysates of I-tic22 (MT, lanes 1, 3, 5) or Anabaena sp. (WT, lanes 2, 4, 6) at indicated protein amounts (mg) with anaTic22 antibodies. Loading was controlled by Ponceau staining. D, 2 μl of indicated strains (2.5 mg chlorophyll ml⁻¹) were spotted on BG11 (panels 1 and 2), BG11₀ (panels 3 and 4), or BG11_NH4 plates (NH₄, panel 5) containing 30 mm lysozyme (panels 2 and 4; images after 7 days). E, the ultrastructure of the outer membrane of the indicated strains was analyzed by electron microscopy as described under “Experimental Procedures.” Additional images are shown in supplemental Fig. S2.