Enzymatic synthesis of a segment of bacteriophage Qβ coat protein gene

Yo Kikuchi, Kenji Sakaguchi

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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%.

Original languageEnglish
Pages (from-to)591-598
Number of pages8
JournalNucleic Acids Research
Volume5
Issue number2
DOIs
Publication statusPublished - 1978 Feb
Externally publishedYes

Fingerprint

Addition method
Bacteriophages
Capsid Proteins
Ligases
Polyribonucleotide Nucleotidyltransferase
RNA
Genes
Synthesis
Gene
Proteins
Protein
Oligoribonucleotides
RNA Phages
Ribonuclease T1
Bacteriophage T4
Oligonucleotides
Alkaline Phosphatase
Phosphatases
Polynucleotides

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Enzymatic synthesis of a segment of bacteriophage Qβ coat protein gene. / Kikuchi, Yo; Sakaguchi, Kenji.

In: Nucleic Acids Research, Vol. 5, No. 2, 02.1978, p. 591-598.

Research output: Contribution to journalArticle

Kikuchi, Yo ; Sakaguchi, Kenji. / Enzymatic synthesis of a segment of bacteriophage Qβ coat protein gene. In: Nucleic Acids Research. 1978 ; Vol. 5, No. 2. pp. 591-598.
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