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Targeted Gene Deletion in Cordyceps militaris Using the Split-Marker Approach
HaiWei Lou1,2, ZhiWei Ye1,2, Fan Yun3
1Department of Bioengineering, College of Food Science and Institute of Food Biotechnology, South China Agricultural University, Guangzhou, 510640, China.
Molecular Biotechnology
|April 2, 2018
Summary
Researchers developed a novel split-marker genetic system for Cordyceps militaris, improving gene knockout efficiency. This method aids in understanding the functions of unknown genes in this valuable medicinal fungus.
Area of Science:
- Mycology
- Molecular Biology
- Genetic Engineering
Background:
- Cordyceps militaris is a macrofungus rich in bioactive compounds regulated by genes.
- The functions of many genes in C. militaris remain uncharacterized.
- Efficient genetic tools are needed to study gene function in C. militaris.
Purpose of the Study:
- To develop a novel genetic transformation system for C. militaris.
- To improve the efficiency of homologous integration for gene knockout.
- To investigate the function of a terpenoid synthase (Tns) gene.
Main Methods:
- Development of a split-marker based genetic system.
- Construction of linear and split-marker deletion cassettes.
- PEG-mediated transformation of C. militaris protoplasts.
- Targeted gene disruption of a Tns gene.
Main Results:
- The split-marker approach significantly enhanced targeted gene disruption efficiency compared to linear deletion cassettes.
- A PEG-mediated protoplast transformation system was successfully established.
- Stable genetic transformants were obtained.
- Tns gene deletion mutants exhibited a distinct color phenotype compared to wild-type.
Conclusions:
- The developed split-marker system is an effective tool for targeted gene deletion in C. militaris.
- This method facilitates the investigation of gene function in C. militaris.
- The study provides a foundation for further genetic studies in this fungus.
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