FIG. 5.
In SW2, foxE, foxY, and foxZ are cotranscribed in the presence of Fe(II). (A) Cloning strategy to identify foxE, foxY, and foxZ, genes from SW2 that enhance light-dependent Fe(II) oxidation activity in SB1003. Numbered arrows identify the positions of the oligonucleotides used for RT-PCR experiments (described in Table 2). The 1-kb scale bar applies to the P3-gm2 segment only. With total RNA extracted from H2-grown, Fe(II)-induced SW2 as a template for cDNA synthesis and oligonucleotides 5 and 6 (B) and 7 and 8 (C), PCR products were obtained for both of the regions between the fox genes (+RT lanes). With total RNA extracted from H2-grown SW2 as a template for cDNA synthesis and oligonucleotides 5 and 6 (D) and 7 and 8 (E), a PCR product was obtained for the region between foxE and foxY but not for that between foxY and foxZ. Additional experiments with oligonucleotides 1, 2, 3, and 4 confirmed the results in panels B, C, D, and E (data not shown). −RT lanes, controls with no reverse transcriptase added to the cDNA reaction mixtures; + lanes, controls with SW2 genomic DNA; − lanes, no-template controls.