Solid-phase synthesis of oligo(2'-deoxyxylonucleotides) and PCR amplification of base-modified DNA fragments.
1-(2'-Deoxy-beta-D-threo-pentofuranosyl)thymine (xTd) and -adenine (xAd) were converted into their appropriately protected 3'-phosphonates 1a, 2a as well as their 2-cyanoethyl phosphoramidites 1b, 2b. These compounds were used for solid-phase syntheses of the oligo(2'-deoxy-beta-D-xylonucleotides) 5-8. Structural properties and behavior against nucleases is described. Apart from oligo(2'-deoxyxylonucleotides) the PCR-amplification of a pUC18 DNA fragment with Taq polymerase was studied in the presence of the 7-deazapurine derivatives of dGTP, dATP, and dITP. The incorporation efficiency of the modified compounds was compared with those of the parent nucleotides. 7-Deaza-2'-deoxyguanosine protected the DNA-fragment from hydrolysis by the restriction endodeoxyribonuclease Eco RI, Pst I, Bam HI, and Sma I if the nucleoside was located within the recognition site.
Seela
F
F
Laboratorium für Organische und Bioorganische Chemie, Fachbereich Biologie/Chemie, Universität Osnabrück, FRG.
Rosemeyer
H
H
Krecmerova
M
M
Röling
A
A
D000241:Adenosine / Q000031:analogs & derivatives; D001483:Base Sequence; D004247:DNA / Q000138:chemical synthesis* / Q000737:chemistry; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D063065:Organophosphonates; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping; D013379:Substrate Specificity; D019914:Taq Polymerase; D013936:Thymidine / Q000031:analogs & derivatives; D014994:Xylose
D009841:Oligonucleotides; D063065:Organophosphonates; C077505:1-(2'-deoxy-beta-threopentofuranosyl)adenine; C077504:1-(2'-deoxy-beta-threopentofuranosyl)thymine; D004247:DNA; D014994:Xylose; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase; D004262:DNA Restriction Enzymes; D000241:Adenosine; D013936:Thymidine