Construction of Escherichia coli amber suppressor tRNA genes. III. Determination of tRNA specificity.
J Mol Biol
; 213(4): 719-26, 1990 Jun 20.
Article
en En
| MEDLINE
| ID: mdl-2141650
ABSTRACT
Using synthetic oligonucleotides, we have constructed a collection of Escherichia coli amber suppressor tRNA genes. In order to determine their specificities, these tRNAs were each used to suppress an amber (UAG) nonsense mutation in the E. coli dihydrofolate reductase gene fol. The mutant proteins were purified and subjected to N-terminal sequence analysis to determine which amino acid had been inserted by the suppressor tRNAs at the position of the amber codon. The suppressors can be classified into three groups on the basis of the protein sequence information. Class I suppressors, tRNA(CUAAla2), tRNA(CUAGly1), tRNA(CUAHisA), tRNA(CUALys) and tRNA(CUAProH), inserted the predicted amino acid. The class II suppressors, tRNA(CUAGluA), tRNA(CUAGly2) and tRNA(CUAIle1) were either partially or predominantly mischarged by the glutamine aminoacyl tRNA synthetase. The class III suppressors, tRNA(CUAArg), tRNA(CUAAspM), tRNA(CUAIle2), tRNA(CUAThr2), tRNA(CUAMet(m)) and tRNA(CUAVal) inserted predominantly lysine.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Supresión Genética
/
Tetrahidrofolato Deshidrogenasa
/
ARN de Transferencia
/
Escherichia coli
Idioma:
En
Revista:
J Mol Biol
Año:
1990
Tipo del documento:
Article