Characterization of the site-specific recombination system of phage ΦD145, and its capacity to promote recombination in human cells.
Phage integrases have the potential of becoming tools for safe site-specific integration of genes into unmodified human genomes. The P2-like phages have been found to have different bacterial host integration sites and consequently they have related integrases with different sequence specificities. In this work the site-specific recombination system of the P2-like phage ΦD145 is characterized. The minimal attB site is determined to 22 nt with 18 nt identity to the core region of attP. A non-coding sequence on the human chromosome 13 is shown to be a rather good substrate for recombination in vivo in bacteria as well as in a plasmid system in HeLa cells when HMG protein recognition sequences are inserted between the left arm-binding site and the core in the complex phage attachment site attP. Thus ΦD145 integrase that belongs to the tyrosine family shows potential as a tool for site-specific integration into the human genome.
Mandali
Sridhar
S
Dept. of Genetics, Microbiology and Toxicology, Stockholm University, Stockholm, Sweden.
Cardoso-Palacios
Carlos
C
Sylwan
Lina
L
Haggård-Ljungquist
Elisabeth
E
D001287:Attachment Sites, Microbiological; D001435:Bacteriophages / Q000201:enzymology*; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D019426:Integrases / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids; D011995:Recombination, Genetic; D017385:Sequence Homology
D019426:Integrases