Genetic transformation of Monascus purpureus DSM1379

Area of Science:

• Microbiology
• Molecular Biology
• Fungal Genetics

Background:

• Monascus purpureus is a fungus with various applications, but efficient genetic transformation methods are limited.
• Developing reliable transformation protocols is crucial for its genetic manipulation and functional studies.

Purpose of the Study:

• To establish an efficient method for the genetic transformation of Monascus purpureus.
• To develop a selectable marker system for Monascus purpureus using hygromycin B resistance.

Main Methods:

• Electroporation was used to introduce the hygromycin B phosphotransferase (hph) gene into Monascus purpureus strains.
• The hph gene was fused to either the Aspergillus nidulans trpC promoter or a Monascus purpureus gpd1 promoter.
• The transformed strain was further modified with the super-folder green fluorescent protein (sgfp) gene.

Main Results:

• Monascus purpureus DSM1397 was identified as a competent recipient strain for transformation.
• Transformed strains exhibited resistance to high concentrations of hygromycin B (up to 500 mg/L) and maintained this resistance for at least five generations.
• Successful transformation was confirmed by the expression of the green fluorescent protein (sgfp) gene, visualized via fluoromicroscopy.

Conclusions:

• A robust hygromycin B resistance-based transformation system for Monascus purpureus has been established.
• This method facilitates the genetic engineering of Monascus purpureus, opening avenues for its improved utilization and research.
• The developed system is suitable for introducing model genes like sgfp, demonstrating its utility for future functional genomics studies.