In vitro selection of RNA aptamer against Escherichia coli release factor 1.
Bioorg Med Chem Lett
; 17(5): 1216-20, 2007 Mar 01.
Article
en En
| MEDLINE
| ID: mdl-17188871
ABSTRACT
A pool of 84-nt RNAs containing a randomized sequence of 50 nt was selected against gel-immobilized Escherichia coli release factor 1 (RF-1) responsible for translation termination at amber (UAG) stop codon. The strongest aptamer (class II-1) obtained from 43 clones bound to RF-1, but not to UAA/UGA-targeting RF-2, with Kd = 30+/-6 nM (SPR). A couple of unpaired hairpin domains in the aptamer were suggested as the sites of attachment of RF-1. By binding to and hence inhibiting the action of RF-1 specifically or bio-orthogonally, aptamer class II-1 enhanced the amber suppression efficiency in the presence of an anticodon-adjusted (CUA) suppressor tRNA without practically damaging the protein translation machinery of the cell-free extract of E. coli, as confirmed by the translation of amber-mutated (gfp(amber141) or gfp(amber178)) and wild-type (gfp(wild)) genes of GFP.
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Base de datos:
MEDLINE
Asunto principal:
Factores de Terminación de Péptidos
/
Proteínas de Escherichia coli
/
Evaluación Preclínica de Medicamentos
/
Aptámeros de Nucleótidos
Tipo de estudio:
Clinical_trials
Idioma:
En
Revista:
Bioorg Med Chem Lett
Asunto de la revista:
BIOQUIMICA
/
QUIMICA
Año:
2007
Tipo del documento:
Article