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Appl Microbiol Biotechnol (2011) 92:227239 DOI 10.1007/s00253-011-3519-5
Received: 10 June 2011 /Revised: 19 July 2011 /Accepted: 25 July 2011 /Published online: 7 August 2011
MINI-REVIEW
Site-specific recombinases as tools for heterologous gene integration
Nobutaka Hirano & Tetsurou Muroi & Hideo Takahashi &
Mitsuru Haruki
# Springer-Verlag 2011
Abstract Site-specific recombinases are the enzymes that catalyze site-specific recombination between two specific DNA sequences to mediate DNA integration, excision, resolution, or inversion and that play a pivotal role in the life cycles of many microorganisms including bacteria and bacteriophages. These enzymes are classified as tyrosine-type or serine-type recombinases based on whether a tyrosine or serine residue mediates catalysis. All known tyrosine-type recombinases catalyze the formation of a Holliday junction intermediate, whereas the catalytic mechanism of all known serine-type recombinases includes the 180 rotation and rejoining of cleaved substrate DNAs. Both recombinase families are further subdivided into two families; the tyrosine-type recombinases are subdivided by the recombination directionality, and the serine-type recombinases are subdivided by the protein size. Over more than two decades, many different site-specific recombinases have been applied to in vivo genome engineering, and some of them have been used successfully to mediate integration, deletion, or inversion in a wide variety of heterologous genomes, including those from bacteria to higher eukaryotes. Here, we review the recombination mechanisms of the best characterized recombinases in each site-specific recombinase family and recent advances in the application of these recombinases to genomic manipulation,
especially manipulations involving site-specific gene integration into heterologous genomes.
Keywords Genome engineering . Integration . Serine-type recombinase . Site-specific recombination . Tyrosine-type recombinase
Introduction
Site-specific recombination systems found in bacteria, bacteriophage, and yeast play a pivotal role in the life cycles of these organisms. These roles include resolution of replicated phage, plasmids, or transposable element DNA (Austin et al. 1981; Broach et al. 1982; Rice et al. 2010), phase variations of bacterial and phage virulence factors (Silverman and Simon 1980; Van de Putte et al. 1980), genomic rearrangements during sporulation or heterocyst differentiation (Sato et al. 1990; Carrasco et al. 1994), and integration of phage genomes into host genomes (Enquist et al. 1979; Groth and Calos 2004). Site-specific recombination is mediated by enzymes called site-specific recombinases, and these enzymes catalyze recombination between two specific DNA sequences to...