Nucleotide sequence of a nicking site of the Streptomyces plasmid pSN22 replicating by the rolling circle mechanism.

Abstract

A putative nicking site in the double strand origin (DSO) of the Streptomyces plasmid pSN22 was identified by comparing the nucleotide sequence of the DSO region with those of two other Streptomyces plasmids, pIJ101 and pJVI. A 7-bp sequence of this putative nicking site, 5'-CTTGGGA-3', was similar to the consensus sequence of the nicking site of the pC194 group of plasmids. When several point mutations were introduced into this 7-bp sequence, the transformation abilities of the mutant plasmid molecules for Streptomyces lividans were either reduced or lost. Southern hybridization analysis indicated that these mutant plasmids could not replicate in S. lividans, but were integrated into the chromosomal DNA.