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

Area of Science:

• Molecular Biology
• Genetics
• Biochemistry

Background:

• Fungal mitochondrial genomes harbor homing endonucleases, enzymes with DNA site-specific cleavage.
• Homing endonucleases are promising tools for genome editing and gene therapy.
• Naturally occurring endonucleases offer an alternative to synthetic enzyme development.

Purpose of the Study:

• To develop methods for identifying homing endonuclease genes in fungal mitochondria.
• To establish protocols for purifying and characterizing these native enzymes.
• To leverage fungal biodiversity for novel genome editing tools.

Main Methods:

• PCR-based screening of fungal mitochondrial rRNA genes.
• Identification of homing endonuclease encoding sequences.
• Purification and biochemical characterization of putative native homing endonucleases.

Main Results:

• Established effective PCR screening methods for fungal mitochondrial DNA.
• Identified potential homing endonuclease sequences within fungal rRNA genes.
• Developed protocols for enzyme purification and functional analysis.

Conclusions:

• Fungal mitochondrial genomes are a viable source for discovering novel homing endonucleases.
• The described methods facilitate the bioprospecting of these enzymes.
• This research provides a foundation for utilizing native fungal endonucleases in biotechnology.