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Nucleic Acids Research, 1978, Vol. 5, No. 2 591-598
© 1978

Articles

Enzymatic synthesis of a segment of bacteriophage Qß coat protein gene

Yo Kikuchi and Kenji Sakaguchi

Laboratory of Microbiological Chemistry, Mitsubishi-Kasei Institute of Life Sciences 11 Minamiooya, Machida, Tokyo 194, Japan

Received January 3, 1978. The oligoribonucleotide, A-A-A-C-U-U-U-Gp, constituting a segment of RNA bacteriophage Qß 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%.


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