In vitro repair of double-strand breaks accompanied by recombination in bacteriophage T7 DNA.
A double-strand break in a bacteriophage T7 genome significantly reduced the ability of that DNA to produce viable phage when the DNA was incubated in an in vitro DNA replication and packaging system. When a homologous piece of T7 DNA (either a restriction fragment or T7 DNA cloned into a plasmid) that was by itself unable to form a complete phage was included in the reaction, the break was repaired to the extent that many more viable phage were produced. Moreover, repair could be completed even when a gap of about 900 nucleotides was put in the genome by two nearby restriction cuts. The repair was accompanied by acquisition of a genetic marker that was present only on the restriction fragment or on the T7 DNA cloned into a plasmid. These data are interpreted in light of the double-strand gap repair mode of recombination.
Masker
W
W
Department of Biochemistry and Molecular Biology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140.
D002474:Cell-Free System; D004249:DNA Damage; D004260:DNA Repair; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004926:Escherichia coli; D066298:In Vitro Techniques; D011995:Recombination, Genetic; D012764:Shigella sonnei; D013604:T-Phages / Q000235:genetics* / Q000378:metabolism; D014771:Virion / Q000254:growth & development
D004262:DNA Restriction Enzymes