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 language | English |
|---|---|
| Pages (from-to) | 591-598 |
| Number of pages | 8 |
| Journal | Nucleic Acids Research |
| Volume | 5 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 1978 Feb |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Enzymatic synthesis of a segment of bacteriophage Qβ coat protein gene
AU - Kikuchi, Yo
AU - Sakaguchi, Kenji
PY - 1978/2
Y1 - 1978/2
N2 - 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%.
AB - 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%.
UR - http://www.scopus.com/inward/record.url?scp=0017843312&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0017843312&partnerID=8YFLogxK
U2 - 10.1093/nar/5.2.591
DO - 10.1093/nar/5.2.591
M3 - Article
C2 - 416426
AN - SCOPUS:0017843312
VL - 5
SP - 591
EP - 598
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 2
ER -