Addition of Short Guanylyl Blocks to Oligonucleotide Primers with a Thermophilic Polynucleotide Phosphorylase
KIKUCHI, Yo; HIRAI, Keiko; SAKAGUCHI, Kenji; Laboratory of Microbiological Chemistry, Mitsubishi-Kasei Institute of Life SciencesMinamiooya, Machida, Tokyo 194
Журнал:
Journal of Biochemistry
Дата:
1979
Аннотация:
Polynucleotide phosphorylase from Thermus thermophilus catalyzed the addition of short guanylyl blocks from GDP to the 3'-hydroxyI termini of oligonucleotide primers at low temperature in a simple reaction mixture. Polyguanylic acid formation was inhibited at 37°C, but the addition of one or two guanylyl residues to oligonucleotide primers proceeded in high yields. The reaction was applied to the synthesis of oligonucleotides containing guanylyl residues at the 3'-end. Using (Ap)<sub>2</sub>A and (Up)<sub>2</sub>U as primers, (Ap)<sub>3</sub>G, (Ap)<sub>3</sub>GpG, and (Up)<sub>2</sub>G were synthesized in yields of 25–52%. (Ap)<sub>2</sub>GpG was synthesized from ApA and GDP in a yield of 13%.
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