Enzymatic synthesis of a segment of bacteriophage Qβ coat protein gene

Yo Kikuchi, Kenji Sakaguchi

研究成果: Article査読

8 被引用数 (Scopus)

抄録

The oligoribonucleotide, A-A-A-C-U-U-U-Gp, constituting a segment of RNA bacteriophage Qβ coat protein gene was efficiently synthesized at a milligram scale by a combination of enzymatic methods using bacteriophage T4 RNA ligase and the thermophilic polynucleotide phosphorylase. A-A-A-Cp was synthesized from A-A-A and pCp by the newly developed mononucleotide addition method using T4 RNA ligase in a yield of 83%, followed by dephosphorylation with bacterial alkaline phosphatase to obtain A-A-A-C. pU-U-U-Gp was synthesized from pU-U-U and GDP by the simultaneous action of polynucleotide phosphorylase and RNase T1 in a yield of 32%. Finaly, the two oligonucleotides (A-A-A-C and pU-U-U-Gp) were ligated with T4 RNA ligase and the octanucleotide, A-A-A-C-U-U-U-Gp, was obtained in a yield of 85%.

本文言語English
ページ(範囲)591-598
ページ数8
ジャーナルNucleic Acids Research
5
2
DOI
出版ステータスPublished - 1978 2
外部発表はい

ASJC Scopus subject areas

  • 統計学、確率および不確実性
  • 応用数学
  • 健康、毒物学および変異誘発
  • 毒物学
  • 遺伝学(臨床)
  • 遺伝学

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