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Cloning an Aspergillus nidulans developmental gene by transformation
The EMBO Journal
|May 1, 1985
Summary
Researchers created a new Aspergillus nidulans transformation system for high-frequency gene cloning. This method enabled the isolation of the developmental gene brlA using visual selection and a specific marker.
Area of Science:
- Molecular Biology
- Mycology
- Genetics
Background:
- Aspergillus nidulans is a model organism for studying fungal development and genetics.
- Efficient transformation systems are crucial for gene cloning and functional analysis in fungi.
Purpose of the Study:
- To develop a high-frequency transformation system for Aspergillus nidulans.
- To isolate and clone the developmental gene brlA using the new system.
Main Methods:
- Development of a transformation vector containing the argB+ selective marker.
- Transformation of Aspergillus nidulans and screening for transformants.
- Isolation and cloning of the brlA gene via visual selection.
- Recovery of integrated plasmids in Escherichia coli.
Main Results:
- Achieved a transformation frequency of 500 stable transformants per microgram of plasmid DNA.
- Successfully isolated and cloned the Aspergillus nidulans developmental gene brlA.
- Evidence suggests transformation occurs via integration, with frequent spontaneous plasmid excision.
Conclusions:
- The developed transformation system is highly efficient for Aspergillus nidulans.
- This system facilitates gene cloning and functional studies, exemplified by the brlA gene isolation.
- The system allows for recovery of integrated plasmids, aiding further genetic manipulation.