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Team:Alberta/References/Publications/Phage integrases: biology and applications
From 2009.igem.org
Amy C. Groth and Michele P. Calos
J. Mol. Biol. (2004) 335, 667–678
Abstract:Phage integrases are enzymes that mediate unidirectional site-specific
recombination between two DNA recognition sequences, the phage
attachment site, attP, and the bacterial attachment site, attB. Integrases
may be grouped into two major families, the tyrosine recombinases and
the serine recombinases, based on their mode of catalysis. Tyrosine family
integrases, such as lambda integrase, utilize a catalytic tyrosine to mediate
strand cleavage, tend to recognize longer attP sequences, and require
other proteins encoded by the phage or the host bacteria. Phage integrases
from the serine family are larger, use a catalytic serine for strand cleavage,
recognize shorter attP sequences, and do not require host cofactors. Phage
integrases mediate efficient site-specific recombination between two
different sequences that are relatively short, yet long enough to be specific
on a genomic scale. These properties give phage integrases growing
importance for the genetic manipulation of living eukaryotic cells,
especially those with large genomes such as mammals and most plants,
for which there are few tools for precise manipulation of the genome. Integrases
of the serine family have been shown to work efficiently in mammalian
cells, mediating efficient integration at introduced att sites or
native sequences that have partial identity to att sites. This reaction has
applications in areas such as gene therapy, construction of transgenic
organisms, and manipulation of cell lines. Directed evolution can be
used to increase further the affinity of an integrase for a particular native
sequence, opening up additional applications for genomic modification.
Link: JMB
