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

摘要

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.