Rapid synthesis of long-chain deoxyribooligonucleotides by the N-methylimidazolide phosphotriester method

Abstract

A modified phosphotriester method has been employed for the efficient chemical synthesis of long-chain deoxyribooligonucleotides. During the course of this work, a general and rapid procedure was developed for the preparation of 24-62-mers in solution. Preparative reversed phase coluimn chromatography on silanized silica gel was used to purify triester intermediates starting from 10-mers. The rapid synthesis of 32-mer and 42-mer on glass and silica gel supports using suitably protected 2-8-mer blocks as coupling units has been also accomplished. In particular, a convenient procedure for the solid-phase synthesis of oligonucleotide blocks bearing 3'-terminal phosphodiester groups is described.

Intracellular precursors and secretion of alkaline extracellular protease of Yarrowia lipolytica.

“…Oligonucleotide probes designed to hybridize to the XPR2 gene were kindly synthesized by B. Dujon using the solid-phase phosphotriester method (21). The probes, 51 and 44 bases in length, were synthesized based on the N-terminal amino acid sequence of mature AEP utilizing the codon bias of the Y. lipolytica LEU2 gene (25).…”

Chemical Synthesis of 80-mer Thymidylic Acid

“…13 The carbon-l.3 spectrum (in pyridine-d s ) of the protected thymidylic acid 8 shows the characteristic signals at 55.3 and 113.8 ppm for DMTr, and that at 31.5 ppm for tert-Bu. Starting from 8, elongation of the oligomer chain was carried out in a similar manner to that already described, the results being summarized in Table II. All the condensation products exhibited 4 both signals of DMTr and tert-Bu in their carbon-13 spectra, although their relative intensity decreased with the increasing number of thymidylic acid units. The purification of higher oligomers than 40-mer was carried out again by a combination of column chromatography on silica gel and gel permeation on Toyopearl HW50F.…”

“…obtained 62-meric oligodeoxyribonucleotide by sequentially adding oligomer blocks of not longer than IO-mer length to the 5 / -terminus. 4 ) Solid-phase approaches have also succeeded in .the synthesis of higher oligomers including 31-mer,5) 42-mer 4 ) and 51-mer. 6 ) However, few papers have discussed the maximum molecular size of.…”

Cyclic Oligoribonucleotides (RNA) by Solid-Phase Synthesis

“…The linear reference sequences rACGU-3'-p and rU 2 GA 4 GAGA 2 C 3 U-3'-p were synthesized on a solid support with a starter group of the type described in ref. [17]. All sequences were synthesized trityl-off.…”