A new assay for tRNA aminoacylation kinetics.
RNA
; 4(8): 1019-23, 1998 Aug.
Article
en En
| MEDLINE
| ID: mdl-9701292
ABSTRACT
An improved quantitative assay for tRNA aminoacylation is presented based on charging of a nicked tRNA followed by separation of an aminoacylated 3'-fragment on an acidic denaturing polyacrylamide gel. Kinetic parameters of tRNA aminoacylation by Escherichia coli AlaRS obtained by the new method are in excellent agreement with those measured by the conventional method. This assay provides several advantages over the traditional methods of measuring tRNA aminoacylation (1) the fraction of aminoacyl-tRNA is measured directly; (2) data can be obtained at saturating amino acid concentrations; and (3) the assay is significantly more sensitive.
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Aminoacil-ARN de Transferencia
/
Alanina-ARNt Ligasa
Idioma:
En
Revista:
RNA
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
1998
Tipo del documento:
Article
País de afiliación:
Estados Unidos