Isolation of high-affinity GTP aptamers from partially structured RNA libraries
作者： J. H. Davis , J. W. Szostak
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摘要: Aptamers, RNA sequences that bind to target ligands, are typically isolated by in vitro selection from libraries containing completely random sequences. To see whether higher-affinity aptamers can be partially structured libraries, we selected for GTP, starting a mixture of fully and libraries. Because stem-loops common motifs previously characterized aptamers, designed the library contain centrally located stable stem-loop. We used an off-rate protocol maximize enrichment high-affinity aptamers. The produced surprisingly large number distinct sequence secondary structures, including seven different with Kds ranging 500 25 nanomolar. engineered stem-loop was present three highest affinity 12 13 independent isolates single consensus sequence, suggesting its inclusion increased abundance pool.
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