Abstract:
:A general approach for the synthesis of oligonucleotide-triplet phosphoramidites and the synthesis of four such blocks are described. A strategy was devised to minimize the number of dimer precursors needed for synthesis of a complete set of triplet-amidite blocks encoding all 20 amino acids. Whereas synthesis of 20 triplet-amidite blocks consisting of codon sequences requires 16 dimer blocks, just seven dimer blocks are required to synthesize all required antisense sequences. The antisense sequences are then converted to codons in template mediated replication. Using a mixture of four triplet-amidites and conventional automated solid-phase DNA synthesis, short (6mer) and medium length (30mer) oligonucleotide mixtures were synthesized and analyzed. The latter was replicated in vitro and used as a mutagenic cassette to produce four mutants of Asp 221 in the enzyme thymidylate synthase. The method establishes the direction and utility for the production and use of triplet-amidite blocks in DNA synthesis.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Ono A,Matsuda A,Zhao J,Santi DVdoi
10.1093/nar/23.22.4677subject
Has Abstractpub_date
1995-11-25 00:00:00pages
4677-82issue
22eissn
0305-1048issn
1362-4962pii
5a0441journal_volume
23pub_type
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