Method of artificial DNA splicing by directed ligation (SDL).
Nucleic Acids Res
; 19(24): 6757-61, 1991 Dec 25.
Article
em En
| MEDLINE
| ID: mdl-1662363
ABSTRACT
An approach to directed genetic recombination in vitro has been devised, which allows for joining together, in a predetermined way, a series of DNA segments to give a precisely spliced polynucleotide sequence (DNA splicing by directed ligation, SDL). The approach makes use of amplification, by means of several polymerase chain reactions (PCR), of a chosen set of DNA segments. Primers for the amplifications contain recognition sites of the class IIS restriction endonucleases, which transform blunt ends of the amplification products into protruding ends of unique primary structures, the ends to be used for joining segments together being mutually complementary. Ligation of the mixture of the segments so synthesized gives the desired sequence in an unambiguous way. The suggested approach has been exemplified by the synthesis of a totally processed (intronless) gene encoding human mature interleukin-1 alpha.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
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DNA Recombinante
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Enzimas de Restrição do DNA
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Interleucina-1
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Genes Sintéticos
Idioma:
En
Revista:
Nucleic Acids Res
Ano de publicação:
1991
Tipo de documento:
Article