Paper Disk를 지지체로 이용한 Oligodeoxyribonucleotides 화학적 합성

Chemical Synthesis of Oligodeoxyribonucleotides Using Paper Disk as a Support

Whatmann chromatography paper(3MM)을 지지체로 사용하여 phosphite-triester법에 의해 미량의 염기가 부착된 5'-d(GAATTCCGGCCA)와 5'-d(GGAGCTGTC)의 두 oligodeoxyribonucleotide를 합성 하였다. Trityl yield 분석에 의하면 76.1${\%}$와 86.5${\%}$의 평균 coupling 수율을 각각 얻었다.두 oligodeoxyribonucleotide들을 HPLC를 이용하여 분리 정제할 수 있었고 snake venom phosphodiesterase와 bacterial alkaline phosphatase에 의해 분리 정량화할 수 있었으며 이때의 염기의 수는 합성을 시도한 염기수와 일치했다. 이처럼 paper를 지지체로 이용한 oligodeoxyribonucleotide의 합성은 값싸고 동시에 많은 염기가 부착된 미량의 DNA를 빠르고 짧은 기간에 수행할 수 있는 장점이 있다.

The two deoxyribonucleotides, 5'-d(GAATTCCGGCCA) and 5'-d(CGAGCTGTC), were synthesized on disks of Whatmann chromatography paper (3MM) by the phosphite-triester method. The average yield of the dodecamer and the nonamer that was measured by trityl analysis is 76.1${\%}$ and 86.5${\%}$ respectively. Separation of the synthesized oligodeoxyribonucleotides were purified by HPLC. The two oligodeoxymers'bases were quantified by HPLC with snake venom phosphodiesterase and bacterial alkaline phosphatase. The numbers of bases were equal to the designed ones. The advantage of using a paper disk as a support allows us to synthesize many different oligodeoxyribenucleotides during one synthesis. This method is the most convenient synthetic technique to synthesize oligodeoxyribonucleotides easily and inexpensively in a relatively short time.