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. 1998 Jul 7;95(14):7969–7974. doi: 10.1073/pnas.95.14.7969

Figure 1.

Figure 1

Trapping the initial hairpin cleavage products generated by SbcCD. (A) An autoradiograph illustrating the DNA fragments produced by SbcCD over a 30-min time period. SbcCD was incubated with 3′-end-labeled hairpin substrate (3′HP78), in the presence of ATP at 37°C, for 0 min (lane b), 1 min (lane c), 5 min (lane d), 10 min (lane e), or 30 min (lane f). 3′HP78 also was incubated for 30 min in the absence of SbcCD (lane a). Early products produced by SbcCD are indicated. (B) Effect of various nucleotides on the SbcCD hairpin-nuclease activity. SbcCD was incubated with 3′-end-labeled hairpin substrate (3′HP78), at 16°C for 30 min, in the presence of various nucleotides. Reactions were performed in the presence of ATP (lane b), dATP (lane c), ADP (lane d), ATPγS (lane e), GTP (lane f), GDP (lane g), or no nucleotide (lane h). 3′HP78 also was incubated with ATP in the absence of SbcCD (lane a). Products were visualized by gel electrophoresis. The sizes of the fragments produced in the presence of ATPγS (lane e) and GTP (lane f) are indicated.