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. © Springer Science+Business Media New York 2014.
CITATION STYLE
Bonocora, R. P., & Belfort, M. (2014). Mapping homing endonuclease cleavage sites using in vitro generated protein. Methods in Molecular Biology, 1123, 55–67. https://doi.org/10.1007/978-1-62703-968-0_4
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