A method for the synthesis of oligonucleotide by single addition of 2'-O-(o-nitrobenzyl)nucleoside 5'-diphosphates using polynucleotide phosphorylase.
Two hexanucleotides A-U-G-U-G-A and C-A-A-U-U-G were synthesized from the chemically synthesized trimers C-A-A and A-U-G by addition of 2'-O-(o-nitrobenzyl)nucleoside diphosphates using polynucleotide phosphorylase isolated from either Escherichia coli or Micrococcus luteus. In each reaction the preference of the enzyme was tested. The o-nitrobenzyl group was removed after addition of the mononucleotide and the deblocked product was isolated by chromatography on DEAE-Sephadex in high yields.
D000244:Adenosine Diphosphate; D001483:Base Sequence; D003565:Cytidine Diphosphate; D004926:Escherichia coli / Q000201:enzymology; D008837:Micrococcus / Q000201:enzymology; D009578:Nitrobenzenes; D009711:Nucleotides; D009841:Oligonucleotides / Q000138:chemical synthesis*; D009843:Oligoribonucleotides / Q000138:chemical synthesis* / Q000302:isolation & purification; D011117:Polyribonucleotide Nucleotidyltransferase; D014530:Uridine Diphosphate
D009578:Nitrobenzenes; D009711:Nucleotides; D009841:Oligonucleotides; D009843:Oligoribonucleotides; D014530:Uridine Diphosphate; D000244:Adenosine Diphosphate; D003565:Cytidine Diphosphate; D011117:Polyribonucleotide Nucleotidyltransferase