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Nucleic Acids Research, 1974, Vol. 1, No. 12 1649-1664
© 1974

Articles

Enzymatic synthesis of deoxyribo-oiigonucleotides of defined sequence. Deoxyribo-oligonucleotide synthesis*

Shirley Gillam, Kimberley Waterman**, Michael Doel*** and Michael Smith

Department of Biochemistry, Faculty of Medicine, University of British Columbia, Vancouver 8, British Columbia, Canada

Received May 30, 1974.

An enzyme, which is probably identical with polynucleotide phosphorylase, was prepared from Escherichia coli B. In thepresence of Mn2$ it catalyzes the addition of one (and to aslight extent more) residue of deoxyribonucleotide residue fromthe diphosphate to an oligodeoxyribonucleotide primer. The shortest effective primers contained three phosphate residues. Ribodinucleotides were effective as primers and accepted twoor three deoxyribonucleotide residues under these conditions. The application of the procedures to the convenient synthesis of certain defined oligodeoxyribonucleotides up to nine residueslong is discussed.

* Research supported by the Medical Research Council of Canada

** Present address: Rockefeller University, New York, N.Y. 10021

*** Medical Research Council of Canada Fellow. Present address: G.D. Searle, Research Division, High Wycombe HP12 4HL, England


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