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J. Biochem, 1976, Vol. 80, No. 1 33-37
© 1976 Japanese Biochemical Society

research-article

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

Yo KIKUCHI, Keiko HIRAI, Kinumi SOMENO and Kenji SAKAGUCHI

Laboratory of Microbial Biochemistry, Mitsubishi-Kasei Institute of Life Sciences Minamiooya, Machida, Tokyo 194

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)2C, (Ap)3U, and (Ap)8G by enzymatic analysis.


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