Efficient suppression of the amber codon in E. coli in vitro translation system.
FEBS Lett
; 579(10): 2156-60, 2005 Apr 11.
Article
em En
| MEDLINE
| ID: mdl-15811334
An mRNA encoding the esterase from Alicyclobacillus acidocaldarius with catalytically essential serine codon (ACG) replaced by an amber (UAG) codon was used to study the suppression in in vitro translation system. Suppression of UAG by tRNA(Ser(CUA)) was monitored by determination of the full-length and active esterase. It was shown that commonly used increase of suppressor tRNA concentration inhibits protein production and therefore limits suppression. In situ deactivation of release factor by specific antibodies leads to efficient suppression already at low suppressor tRNA concentration and allows an in vitro synthesis of fully active enzyme in high yield undistinguishable from wild-type protein.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Biossíntese de Proteínas
/
Genes Supressores
/
Códon de Terminação
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Escherichia coli
Idioma:
En
Ano de publicação:
2005
Tipo de documento:
Article