Preparation of 35S-labeled polyphosphorothioate oligodeoxyribonucleotides by use of hydrogen phosphonate chemistry.
The title compounds were chemically synthesized as their 5'-dimethoxytrityl derivatives by base-catalyzed reaction of 35S-enriched elemental sulfur with support-bound hydrogen phosphonate oligomer. This was derived from adamantane carbonyl chloride-activated coupling of nucleotide hydrogen phosphonate monomers, and similarly activated capping with isopropyl phosphite. A convenient, disposable, reversed-phase cartridge was utilized to purify and isolate the 5'-dimethoxytrityl derivative for subsequent in situ detritylation and elution of the final product. The specific activity obtained for the title compounds was ca. 10(7) cpm/mumols-eq P(O)S-. The procedure should be readily adaptable to appropriate syntheses of other P-S containing analogs of DNA and RNA.
Stein
A
A
Medicine Branch, National Cancer Institute, Bethesda, Maryland 20892.
Iversen
P L
PL
Subasinghe
C
C
Cohen
J S
JS
Stec
W J
WJ
Zon
G
G
D001483:Base Sequence; D004247:DNA / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006678:HIV / Q000032:analysis; D007202:Indicators and Reagents; D007553:Isotope Labeling; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis*; D063065:Organophosphonates / Q000032:analysis*; D012313:RNA / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D013455:Sulfur / Q000032:analysis; D013462:Sulfur Radioisotopes; D013873:Thionucleotides / Q000138:chemical synthesis*
D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D063065:Organophosphonates; D012333:RNA, Messenger; D012367:RNA, Viral; D013462:Sulfur Radioisotopes; D013873:Thionucleotides; D012313:RNA; D013455:Sulfur; D004247:DNA