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Figure 4.
Binding of cry3 to DNA probes containing a single T<>T dimer
in the central position. (A) Sequences and structures of probes.
The T<>T dimer is positioned within the VspI recognition site
(boxed in probe 1). Probe 1 forms a perfect duplex. In probes 2
and 3, the 5′ and 3′ thymines, respectively, of the T<>T
dimer are not hydrogen bonded to the complementary strand. In
probe 3, only one hydrogen bond is formed between the 5′
thymine of the T<>T dimer and the complementary adenine (23). In
probes 4–8, the T<>T lesion is positioned in the center of
loop structures with 2–10 base pairs. Hydrogen bonds between
complementary bases are shown as dashed lines. The upper strand
(50 nt) was labeled at the 5′ position with IRDye700 (MWG
Biotech AG) (marked with asterisk). (B and C) EMSA showing cry3
binding to probes with (B) or without (C) the central T<>T
dimer. Probes shown in A and the single-stranded control (probe
9) were mixed with cry3 (+) or with the same aliquot of buffer
(−). Arrows indicate the positions of shifted bands.
Representative gels from 2 independent experiments are shown.
(D) Quantitative binding data. Mean values and standard errors
of the 2 independent experiments are shown.
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