A 2.2-kilobase repeated DNA segment is associated with DNA amplification in Streptomyces fradiae

Abstract

We have previously identified a 10.5-kilobase DNA sequence which is highly amplified and tandemly repeated in the mutant Streptomyces fradiae JS85. A library of DNA was prepared from S. fradiae T776, which does not contain amplified DNA. The library was screened by plaque hybridization to identify phage clones containing the unamplified 10.5-kilobase DNA sequence. Four phage isolates were identified which contained DNA homology to the amplified DNA sequence. This sequence was designated the amplifiable unit of DNA. None of the clones carried an entire amplifiable unit of DNA, and so overlapping regions were aligned to create a map of the entire region. Detailed restriction mapping identified a 2.2-kilobase direct repeat at the ends of the amplifiable unit of DNA. Analysis by Southern hybridization confirmed that the direct repeats were homologous to each other. The DNA of S. fradiae contained at least two additional copies of DNA that was homologous to the repeat sequence.