New effective method for the synthesis of oligonucleotides via phosphotriester intermediates.
نویسندگان
چکیده
A rapid and convenient method for the synthesis of deoxyribooligonucleotides has been developed using the phosphotriester approach. The advantage of this methodology for work in solution was successfully demonstrated in synthesis of a number of DNA fragments up to 32-long. Adaptation of the presented method to solid-phase synthesis allows a pentadecamer to be assembled in 4-5 hours using dinucleotides as coupling units.
منابع مشابه
Polymer supported synthesis of oligonucleotides by a phosphotriester method.
A new polymer supported synthesis of deoxyribooligonucleotides which can be adapted to automation is described. The method is based on the elongation of an oligomer chain in the 5'- to 3' -end direction using the modified phosphotriester chemistry. The approach is exemplified by the synthesis of a nonanucleotide d(TTCGTCTTG).
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Evidence will be presented to show that the allyl group is suitable for the protection of a 3'-terminal phosphodiester function. The latter will be demonstrated by the synthesis, via a phosphotriester approach, of two cyclic tetraribonucleotides [r(AAAA) and r(UAMe2UAMe2)], two cyclic hexadeoxyribonucleotides [d(CGCGCG) and d(TAAAAA)] and a cyclic octadeoxyribonucleotide [d(CGTGCGTG)].
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A phosphotriester solid phase method on a polyamide support has been used to prepare oligodeoxyribonucleotides up to 12 units long. Compared to solid phase phosphodiester synthesis the new methodology is quicker, more flexible and gives 10-60-fold better overall yields.
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Synthesis of oligonucleotides is turning into a large scale production, since they are increasingly demanded as new powerful drugs, as well as probes and primers in diagnostic kits. Synthesis of large quantities of these nucleic acid derivatives is a prerequisite for their successful commercialization, especially as antisense drugs, which represent their widest field of application. A simple an...
متن کاملChemical synthesis and sequence studies of deoxyribooligonucleotides which constitute the duplex sequence of the lactose operator of Escherichia coli.
We have synthesized the deoxyribooligonucleotide fragments, constituting the sequence of the lac operator of Escherichia coli. Two of these fragments, d(pApApTpTpGpTpTpApT) (nonamer) and d(pApApTpTpGpTpGpApG) (nonamer), corresponding to the 5' termini of lac operator have been synthesized by the phosphodiester method. The remaining four fragments, d(ApCpApApTpT) (hexamer), d(ApTpApApCpApApTpT) ...
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عنوان ژورنال:
- Nucleic acids research
دوره 10 21 شماره
صفحات -
تاریخ انتشار 1982