title: PCR-based bioprospecting for homing endonucleases in fungal mitochondrial rRNA genes.
abstract: Fungal mitochondrial genomes act as "reservoirs" for homing endonucleases. These enzymes with their DNA site-specific cleavage activities are attractive tools for genome editing and gene therapy applications. Bioprospecting and characterization of naturally occurring homing endonucleases offers an alternative to synthesizing artificial endonucleases. Here, we describe methods for PCR-based screening of fungal mitochondrial rRNA genes for homing endonuclease encoding sequences, and we also provide protocols for the purification and biochemical characterization of putative native homing endonucleases.
authors: Department of Biochemistry, Université de Montréal, Montréal, QC, Canada.
authors: Mohamed
authors: M
authors: Hafez
authors: Tuhin Kumar
authors: TK
authors: Guha
authors: Chen
authors: C
authors: Shen
authors: Jyothi
authors: J
authors: Sethuraman
authors: Georg
authors: G
authors: Hausner
mesh_terms: D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D015870:Gene Expression; D018390:Gene Targeting; D005800:Genes, Fungal; D050259:Genes, Mitochondrial; D020459:Genes, rRNA; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA
chemical_list: D004720:Endonucleases