Bacteriophage phiC31 integrase mediated transgenesis in Xenopus laevis for protein expression at endogenous levels.
Bacteriophage phiC31 inserts its genome into that of its host bacterium via the integrase enzyme which catalyzes recombination between a phage attachment site (attP) and a bacterial attachment site (attB). Integrase requires no accessory factors, has a high efficiency of recombination, and does not need perfect sequence fidelity for recognition and recombination between these attachment sites. These imperfect attachment sites, or pseudo-attachment sites, are present in many organisms and have been used to insert transgenes in a variety of species. Here we describe the phiC31 integrase approach to make transgenic Xenopus laevis embryos.
PubMed ID: 19085128
PMC ID: PMC3071032
Article link: Methods Mol Biol.
Grant support: DC007481 NIDCD NIH HHS , GM069944 NIGMS NIH HHS , R01 GM069944-05A2 NIGMS NIH HHS , R01 GM069944 NIGMS NIH HHS