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PCR-based bioprospecting for homing endonucleases in fungal mitochondrial rRNA genes.

Authors
  • Hafez, Mohamed
  • Guha, Tuhin Kumar
  • Shen, Chen
  • Sethuraman, Jyothi
  • Hausner, Georg
Type
Published Article
Journal
Methods in Molecular Biology
Publication Date
Jan 01, 2014
Volume
1123
Pages
37–53
Identifiers
DOI: 10.1007/978-1-62703-968-0_3
PMID: 24510258
Source
Medline
License
Unknown

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.

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