Site-specific recombination in Schizosaccharomyces pombe and systematic assembly of a 400kb transgene array in mammalian cells using the integrase of Streptomyces phage ϕBT1

We have established the integrase of the Streptomyces phage ϕBT1 as a tool for eukaryotic genome manipulation. We show that the ϕBT1 integrase promotes efficient reciprocal and conservative site-specific recombination in vertebrate cells and in Schizosaccharomyces pombe, thus establishing the utility of this protein for genome manipulation in a wide range of eukaryotes. We show that the ϕBT1 integrase can be used in conjunction with Cre recombinase to promote the iterative integration of transgenic DNA. We describe five cycles of iterative integration of a candidate mouse centromeric sequence 80 kb in length into a human mini-chromosome within a human-Chinese hamster hybrid cell line. These results establish the generality of the iterative site-specific integration technique.

• Publication year

  2007

• Venue

  Nucleic Acids Research

• Publication date

  2007-12-20

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1093/nar/gkm1123PMID 18096621PMCID 2248738
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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