title: Mapping homing endonuclease cleavage sites using in vitro generated protein.
abstract: Mapping the precise position of endonucleolytic cleavage sites is a fundamental experimental technique used to describe the function of a homing endonuclease. However, these proteins are often recalcitrant to cloning and over-expression in biological systems because of toxicity induced by spurious DNA cleavage events. In this chapter we outline the steps to successfully express a homing endonuclease in vitro and use this product in nucleotide-resolution cleavage assays.
authors: New York State Department of Health, Wadsworth Center, Albany, NY, USA.
authors: Richard P
authors: RP
authors: Bonocora
authors: Marlene
authors: M
authors: Belfort
mesh_terms: D001665:Binding Sites; D053837:DNA Cleavage; D004720:Endonucleases / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D004720:Endonucleases