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. 2006 Aug 31;34(16):4537–4545. doi: 10.1093/nar/gkl634

Figure 2.

Figure 2

RFLP analysis of chloroplast transformants obtained with rps18 knockout constructs. Equal amounts of extracted total cellular DNA (5 µg per sample) were digested with the restriction enzymes indicated, separated in 0.8% agarose gels, blotted and hybridized to a radiolabeled restriction fragment derived from cloned tobacco plastid DNA (NdeI/SpeI fragment; Figure 1). (A) RFLP with AccI. (B) RFLP with EcoRV and SalI. The probe detects, in addition to the expected fragments (Figure 1), an unexpected band in most samples which is the product of flip-flop recombination (see text for details and Figures 3 and 4). Fragment sizes of the molecular weight marker are given in kb. Lane Nt-Δrps18B-43 is positive in the AccI RFLP and negative in the EcoRV/SalI RFLP most probably because the leaf sample harvested for DNA extraction for the AccI blot was heteroplasmic, whereas the sample harvested for isolating the DNA for the EcoRV/SalI blot had segregated into virtual homoplasmy for the wild-type genome. Wt: wild type.