Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase.

Science (1990), Volume 249, Page 505

Abstract:

High-affinity nucleic acid ligands for a protein were isolated by a procedure that depends on alternate cycles of ligand selection from pools of variant sequences and amplification of the bound species. Multiple rounds exponentially enrich the population for the highest affinity species that can be clonally isolated and characterized. In particular one eight-base region of an RNA that interacts with the T4 DNA polymerase was chosen and randomized. Two different sequences were selected by this procedure from the calculated pool of 65,536 species. One is the wild-type sequence found in the bacteriophage mRNA; one is varied from wild type at four positions. The binding constants of these two RNA's to T4 DNA polymerase are equivalent. These protocols with minimal modification can yield high-affinity ligands for any protein that binds nucleic acids as part of its function; high-affinity ligands could conceivably be developed for any target molecule.

Polymerases:

T4

Topics:

Mutational Analysis, Biotech Applications, Kinetic Parameters, Accessory Proteins/Complexes, Methods

One line summary:

High affinity mRNA sequences were isolated and amplified, both a wild type sequence and a sequence varied from the wild-type in four positions. This procedure can be used to isolate nucleic acids that bind to protein of interest.

Status:

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Results:

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