title: A novel site-specific recombination system derived from bacteriophage phiMR11.
abstract: We report identification of a novel site-specific DNA recombination system that functions in both in vivo and in vitro, derived from lysogenic Staphylococcus aureus phage phiMR11. In silico analysis of the phiMR11 genome indicated orf1 as a putative integrase gene. Phage and bacterial attachment sites (attP and attB, respectively) and attachment junctions were determined and their nucleotide sequences decoded. Sequences of attP and attB were mostly different to each other except for a two bp common core that was the crossover point. We found several inverted repeats adjacent to the core sequence of attP as potential protein binding sites. The precise and efficient integration properties of phiMR11 integrase were shown on attP and attB in Escherichia coli and the minimum size of attP was found to be 34bp. In in vitro assays using crude or purified integrase, only buffer and substrate DNAs were required for the recombination reaction, indicating that other bacterially encoded factors are not essential for activity.
authors.affiliation: Department of Microbiology and Infection, Kochi Medical School, Kohasu Oko-cho, Nankoku, Kochi 783-8505, Japan.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Rashel
authors.forename: Jumpei
authors.initials: J
authors.lastname: Uchiyama
authors.forename: Takako
authors.initials: T
authors.lastname: Ujihara
authors.forename: Iyo
authors.initials: I
authors.lastname: Takemura
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hoshiba
authors.forename: Shigenobu
authors.initials: S
authors.lastname: Matsuzaki
mesh_terms: D001435:Bacteriophages / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D016297:Mutagenesis, Site-Directed / Q000379:methods* / Q000706:statistics & numerical data*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics*; D013211:Staphylococcus aureus / Q000235:genetics / Q000821:virology
chemical_list: D011994:Recombinant Proteins