Synthesis of complementary DNA fragments via phosphotriester intermediates
- 1 January 1979
- journal article
- research article
- Published by Wiley in Recueil des Travaux Chimiques des Pays-Bas
- Vol. 98 (11) , 537-548
- https://doi.org/10.1002/recl.19790981105
Abstract
The preparation of the complementary oligodeoxyribonucleotides Ia,b and II via a modified phosphotriester method is described. The synthesis of these compounds was accomplished by using: (a) the levulinyl group for temporary and persistent blocking of the 5′‐hydroxyl; (b) the monofunctional phosphorylating agent 2,2,2‐trichloroethyl 2‐chlorophenyl phosphorochloridate (3) to introduce the phosphate moiety at the 3′‐position; (c) the benzoyl group to protect the terminal 3′‐hydroxyl; (d) acyl groups for the protection of the exocyclic amino functions of the bases; and (e) 1‐(2,4,6‐triisopropylbenzenesulphonyl)‐3‐nitro‐1,2,4‐triazole (TPS‐NT, 11c) as activating agent in the formation of phosphotriester linkages. Final removal of all protecting groups was performed by fluoride ion treatment followed by aqueous ammonia. The oligonucleotides Ia,b and II were, after purification, isolated in good yields and, as followed from HPLC analysis, in high purity.Keywords
This publication has 43 references indexed in Scilit:
- 4-Nitrophenyl phenyl phosphorochloridate: a new phosphorylating agent for oligonucleotide synthesisJournal of the Chemical Society, Chemical Communications, 1977
- The phosphotriester approach to oligonucleotide synthesis: preparation of oligo- and poly-thymidylic acidsJournal of the Chemical Society, Perkin Transactions 1, 1977
- PhenylN-Phenylphosphoroamidochloridate: A New Phosphorylating AgentSynthesis, 1976
- Nucleotide, II. Über die Verseifung von Dinucleosidmonophosphorsäurephenylester und die Isomerisierung der Internucleotid‐BindungEuropean Journal of Inorganic Chemistry, 1975
- Novel condensing reagents for polynucleotide synthesisJournal of the Chemical Society, Chemical Communications, 1974
- CIV. Total synthesis of the structural gene for an alanine transfer ribonucleic acid from yeast. Chemical synthesis of an icosadeoxynucleotide corresponding to the nucleotide sequence 21 to 40Journal of Molecular Biology, 1972
- The synthesis of oligoribonucleotides. Part IX. Preparation of ribonucleoside 2′-acetal 5′-estersJournal of the Chemical Society C: Organic, 1971
- Studies on Polynucleotides. LII.1 The Use of 2,4,6-Triisopropylbenzenesulfonyl Chloride for the Synthesis of Internucleotide Bonds2Journal of the American Chemical Society, 1966
- Studies on Polynucleotides. XXIV.1 The Stepwise Synthesis of Specific Deoxyribopolynucleotides (4).2 Protected Derivatives of Deoxyribonucleosides and New Syntheses of Deoxyribonucleoside-3″ Phosphates3Journal of the American Chemical Society, 1963
- Studies on Polynucleotides. IX.1 Experiments on the Polymerization of Mononucleotides. Certain Protected Derivatives of Deoxycytidine-5' Phosphate and the Synthesis of Deoxycytidine Polynucleotides2Journal of the American Chemical Society, 1961