Method of artificial DNA spling by directed ligation(SDL)

E. N. Lebedenko, K. R. Birikh, O. V. Plutalov, Yu A. Berlin

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)

Abstract

An approach to directed genetic recombinationin 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-1α.

Original languageEnglish
Pages (from-to)6757-6761
Number of pages5
JournalNucleic Acids Research
Volume19
Issue number24
DOIs
Publication statusPublished - 25 Dec 1991
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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