Journal ArticleDOI
Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase
Craig Tuerk,Larry Gold +1 more
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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.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.read more
Citations
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Light-directed, spatially addressable parallel chemical synthesis.
TL;DR: High-density arrays formed by light-directed synthesis are potentially rich sources of chemical diversity for discovering new ligands that bind to biological receptors and for elucidating principles governing molecular interactions.
Journal ArticleDOI
Rapid evolution of a protein in vitro by DNA shuffling.
TL;DR: It is reported here that selected mutants had a minimum inhibitory concentration of 640 μg ml-1, a 32,000-fold increase and 64-fold greater than any published TEM-1 derived enzyme.
Journal ArticleDOI
Selection of single-stranded DNA molecules that bind and inhibit human thrombin.
TL;DR: The isolation of single-stranded DNA aptamers to the protease thrombin of the blood coagulation cascade is described and binding affinities in the range 25–200 nM are reported.
Journal ArticleDOI
Nanostructured plasmonic sensors.
Matthew E. Stewart,Christopher R. Anderton,Lucas B. Thompson,Joana Maria,Stephen K. Gray,John A. Rogers,Ralph G. Nuzzo +6 more
TL;DR: This work has shown that coherent oscillations of conduction electrons on a metal surface excited by electromagnetic radiation at a metal -dielectric interface can be associated with surface plasmons, which have potential applications in miniaturized optical devices, sensors, and photonic circuits.
Journal ArticleDOI
Listening to silence and understanding nonsense: exonic mutations that affect splicing
TL;DR: As the splicing mechanisms that depend on exonic signals are elucidated, new therapeutic approaches to treating certain genetic diseases can begin to be explored.
References
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Book
Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal ArticleDOI
Compilation and analysis of Escherichia coli promoter DNA sequences.
TL;DR: A consensus promoter sequence based on homologies among 112 well-defined promoters was determined that was in substantial agreement with previous compilations.
Journal ArticleDOI
Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates
TL;DR: A method is described to synthesize small RNAs of defined length and sequence using T7 RNA polymerase and templates of synthetic DNA which contain the T7 promoter to increase the variety of RNAs which can be made.
Journal ArticleDOI
Selection in vitro of an RNA enzyme that specifically cleaves single-stranded DNA
TL;DR: The selected molecule represents the discovery of the first RNA enzyme known to cleave single-stranded DNA specifically and allows evolution experiments to be carried out in response to artificially imposed selection constraints.
Journal ArticleDOI
Isothermal, in vitro amplification of nucleic acids by a multienzyme reaction modeled after retroviral replication.
John C. Guatelli,Kristina Marie Whitfield,Deborah Y. Kwoh,Kevin J. Barringer,Douglas D. Richman,Thomas R. Gingeras +5 more
TL;DR: A target nucleic acid sequence can be replicated exponentially in vitro under isothermal conditions by using three enzymatic activities essential to retroviral replication: reverse transcriptase, RNase H, and a DNA-dependent RNA polymerase, and this reaction accumulates cDNA and RNA copies of the original target.