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The synthesis of blocked triplet-phosphoramidites and their use in mutagenesis.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Ono, A. Matsuda, Akira Zhao, Jindong Santi, Daniel V. |
| Copyright Year | 1995 |
| 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. |
| Starting Page | 1505 |
| Ending Page | 1505 |
| Page Count | 1 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://oup.silverchair-cdn.com/oup/backfile/Content_public/Journal/nar/23/22/10.1093/nar/23.22.4677/2/23-22-4677.pdf?Expires=1492599113&Key-Pair-Id=APKAIUCZBIA4LVPAVW3Q&Signature=ETgzNvtpjQ1Je6MfUPvEQHcMFFoAXkyoNEq92pgO2RWHMW9~6Ac~HpXOfWBn4QxnLAF2XJCHv2rSZzF~MqARztw2LN9CBAYZ9CNgjCgirvMVbUEVVk6-flu8~QuVIDi36KoDxTf5sOA9dTlW-PgzjrHP4tmpM7AU4ZlTkvbhVI3WKkbQhscsyCLkfP9rWOk9KdZfMjqU9-PvJTqCq5Cx~tztPUtrsuSQLdJIkDChSjcuLZNM48Mrifg~64OBJJ3qVO1WKZ4hlNWGPYqa~xW389CAYid49tp5fnmSacdxvBhTNDhtLii35hchCqxdThw4Syw4AZnBZDFiWmJhhsnHLQ__ |
| PubMed reference number | 8524660v1 |
| Volume Number | 23 |
| Issue Number | 22 |
| Journal | Nucleic acids research |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Amino Acids Aspartate Clinical Use Template Codon (nucleotide sequence) DNA Replication Dinucleoside Phosphates License Nucleic Acid Strand Oligonucleotide Synthesis Oligonucleotides Reagents Thymidylate Synthase Trinucleotide Repeat Expansion Trinucleotide Repeats antisense therapy mixture mutant |
| Content Type | Text |
| Resource Type | Article |
