Continuous In Vitro Evolution of a Ribozyme that Catalyzes Three Successive Nucleotidyl Addition Reactions
| Autor: | Gerald F. Joyce, Martin C. Wright, Kathleen E. McGinness |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Stereochemistry
Population Molecular Sequence Data Clinical Biochemistry Biology Polymerase Chain Reaction Biochemistry Catalysis Substrate Specificity Evolution Molecular Viral Proteins Drug Discovery Nucleotide RNA Catalytic education Promoter Regions Genetic Molecular Biology Ligase ribozyme chemistry.chemical_classification Pharmacology education.field_of_study Addition reaction Base Sequence Oligonucleotide Nucleotides Sequence Analysis RNA Ribozyme RNA RNA Ligase (ATP) General Medicine DNA-Directed RNA Polymerases Kinetics chemistry Mutagenesis biology.protein Nucleic Acid Conformation Molecular Medicine Phosphorus Radioisotopes Systematic evolution of ligands by exponential enrichment |
| Zdroj: | Chemistry & Biology. 9(5):585-596 |
| ISSN: | 1074-5521 |
| DOI: | 10.1016/s1074-5521(02)00136-9 |
| Popis: | Variants of the class I ligase ribozyme, which catalyzes joining of the 3' end of a template bound oligonucleotide to its own 5' end, have been made to evolve in a continuous manner by a simple serial transfer procedure that can be carried out indefinitely. This process was expanded to allow the evolution of ribozymes that catalyze three successive nucleotidyl addition reactions, two template-directed mononucleotide additions followed by RNA ligation. During the development of this behavior, a population of ribozymes was maintained against an overall dilution of more than 10(exp 406). The resulting ribozymes were capable of catalyzing the three-step reaction pathway, with nucleotide addition occurring in either a 5' yieldig 3' or a 3' yielding 5' direction. This purely chemical system provides a functional model of a multi-step reaction pathway that is undergoing Darwinian evolution. |
| Databáze: | OpenAIRE |
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