Enzymatic synthesis of oligonucleotides of defined sequence. The "single addition" of 2(3)-O-dihydrocinnamoyl-nucleoside 5'-diphosphate to a primer oligonucleotide catalyzed by a thermophilic polynucleotide phosphorylase.
Several oligonucleotides of defined sequence were synthesized using 2'(3')-O-dihydrocinnamoyl-nucleoside 5'-diphosphates (DHC-NDP) as substrates for polynucleotide phosphorylase [EC 2.7.7.8] from Thermus thermophilus. The enzyme catalyzed the transfer of one nucleotidyl residue from each of the 2'(3')-O-dihydrocinnamoyl esters of CDP, UDP, and GDP to the 3'-terminus of the primer triadenosine diphosphate, (Ap)2A. The products were shown to be (Ap)3C, (Ap)3U, and (Ap)3G by enzymatic analysis.
D000227:Adenine Nucleotides / Q000378:metabolism; D002934:Cinnamates / Q000378:metabolism*; D003597:Cytosine Nucleotides / Q000378:metabolism; D006088:Gram-Negative Aerobic Bacteria / Q000201:enzymology*; D006150:Guanine Nucleotides / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D009841:Oligonucleotides / Q000096:biosynthesis*; D011117:Polyribonucleotide Nucleotidyltransferase / Q000378:metabolism*; D014500:Uracil Nucleotides / Q000378:metabolism
D000227:Adenine Nucleotides; D002934:Cinnamates; D003597:Cytosine Nucleotides; D006150:Guanine Nucleotides; D009841:Oligonucleotides; D014500:Uracil Nucleotides; D008345:Manganese; D011117:Polyribonucleotide Nucleotidyltransferase; D008274:Magnesium