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. 2003 Dec 15;31(24):7208–7215. doi: 10.1093/nar/gkg945

Figure 4.

Figure 4

A chloramphenicol-dependent, UV-induced modification in H.halobium 23S rRNA. (A) Complexes were formed between ribosomes (0.15 µM) and chloramphenicol (1.2 mM) by incubating at 37°C for 20 min. The samples were irradiated for 30 min at 365 nm, followed by isolation of rRNA and primer extension with primer Hh2133. (B) Ribosomes (0.5 µM) were incubated with (+) and without (–) chloramphenicol (1.2 mM) for 20 min at 37°C, irradiated at 365 nm for 30 min, followed by isolation of rRNA. A small portion of each sample (70S) was analyzed directly by extension from primer Hh2133. The remainder of each sample was treated with RNase H in the presence of oligonucleotides Hh1928 and Hh2255. The resulting fragment (Frag.) was gel-purified and subjected to primer extension using primer Hh2133. The rRNA modification-induced stops are displaced by one base below the corresponding stop in the sequencing tracks (G, A, U and C).