Abstract
We have established the integrase of the Streptomyces phage phi BT1 as a tool for eukaryotic genome manipulation. We show that the phi BT1 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 phi BT1 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.
Original language | English |
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Article number | e9 |
Journal | Nucleic Acids Research |
Volume | 36 |
Issue number | 1 |
Early online date | 20 Dec 2007 |
DOIs | |
Publication status | Published - Jan 2008 |
Keywords
- human Y-chromosome
- PHI-C31 integrase
- CRE recombinase
- gene
- efficient
- genome
- DNA
- expression
- centromere
- selection