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Single-strand nicks caused by DNase I are capable of inactivatingB. subtilis transforming DNA. Under suitable conditions, the polynucleotide joining enzyme fromE. coli and a similar DPN-requiring activity fromB. subtilis can completely repair this damage, restore biological activity, and increase the single-strand molecular weight. The rates of inactivation of a single genetic marker and of a four-marker linkage group suggest that a single-strand nick is inactivating even when far from the site of genetic damage.