酵素を用いたオリゴヌクレオチドの合成と応用<ミニノート>

概要

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A new rapid method to synthesize an oligonucleotide 9-mer of any base sequence is proposed. According to the base sequence designed for the oligomer, a 5'tetramer and a 3'tetramer are picked up from a library of tetradeoxyribonucleotides, which have been previously prepared using a DNA synthesizer. The 5'tetramer is added with a mono-ribo-nucleotide (according to the 5th base of the desired sequence) using the terminal deoxynucleotidyl transferase and the 3'tetramer is phosphorylated at 5'terminus using the T4 polynucleotide kinase. Then, the 5'pentamer and the 3'tetramer are combined with the T4 RNA ligase, resulting in a 9-mer having the desired sequence. The 9-mer is proved to serve as a primer for a DNA polymerase reaction without purification steps. This procedure which requires only 2h, can rapidly provide primers for some method of the large-scale DNAsequencmg.
埼玉大学分析センターの論文
1991-11-00