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. 2005 Sep;71(9):5297–5303. doi: 10.1128/AEM.71.9.5297-5303.2005

FIG. 3.

FIG. 3.

Differential expression of the kdp operons in Anabaena sp. strain L-31. (A) Northern blotting. Total RNA was isolated from Anabaena sp. strain L-31 cells grown in BG-11/K5 or BG-11/K0 for 18 and 36 h and electrophoretically resolved (15 μg per lane) on formaldehyde agarose gels. First, 600 bp of kdpA1 ORF, 1 to 400 bp of kdpA2 ORF, and 200 to 600 bp of kdpC2 ORF were amplified by PCR, labeled with DIG, and used as kdpA1, kdpA2, or kdpC2 probes, respectively. The RNA was transferred to nylon membrane and subsequently hybridized to the individual probes. The 5.3-kb transcript is indicated by an arrow. Equal loading of RNA was confirmed by ethidium bromide staining of rRNA from various samples. (B) RT-PCR. Total RNA was isolated from cells grown in BG-11/K0 for 4 or 16 h after inoculation and subjected to RT reaction with primers specific for Anabaena sp. strain L-31 kdpA1, kdpA2, or kdpC1 genes. The RT reaction was subsequently subjected to PCR, and the amplification products were resolved by electrophoresis and stained with ethidium bromide.