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Nucleic Acids Research, 1984, Vol. 12, No. 5 2407-2419
© 1984

MOLECULAR BIOLOGY

In vitro generation of specific deletions in DNA cloned In M13 vectors using synthetic oligodeoxyribonucleotides: mutants in the 5'-flanking region of the yeast alcohol dehydrogenase II gene

Voon-Loong Chan1 and Michael Smith

Department of Biochemistry, University of British Columbia, Vancouver British Columbia V6T 1W5, Canada

Received June 13, 1983. Accepted February 13, 1984.

Deletion mutants are particularly useful in defining the boundaries of noncoding genetic functions. Such mutants can be precisely generated using synthetic oligodeoxyribonucleotides as mutagens. In this paper we describe the application of this method to recombinant DNA cloned in a phage M13-derived vector. The mutagenic oligodeoxyribonucleotides, 20 and 21 nucleotides in length, were used to delete a tract of 20 dA-dT base-pairs and an adjacent 22 base-pair perfect dyad from the ADR3 locus, the 5'-flanking regulatory region of the ADR2 gene, of Saccharomyces cerevisiae with high efficiency.

1Permanent address: Department of Microbiology, University of Toronto, Ontario M5S 1AI, Canada


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