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Cloning of a new bidirectionally selectable marker for Aspergillus strains
F P Buxton1, D I Gwynne, R W Davies
1Allelix Biopharmaceuticals, Mississauga, Ontario, Canada.
Gene
|December 14, 1989
Summary
Researchers cloned the adenosine triphosphate sulfurylase (ATPsase) gene in Aspergillus. This breakthrough enables efficient genetic engineering in industrial Aspergillus strains by selecting for or against ATPsase activity.
Area of Science:
- Molecular Biology
- Mycology
- Biotechnology
Background:
- Mutants lacking adenosine triphosphate sulfurylase (ATPsase) cannot utilize sulfate as a sulfur source and exhibit selenate resistance.
- These ATPsase-deficient mutants (sC-) are easily generated in Aspergillus niger and Aspergillus nidulans via selenate resistance selection.
Purpose of the Study:
- To clone the gene encoding ATPsase from Aspergillus nidulans.
- To demonstrate the utility of the cloned gene for genetic manipulation in Aspergillus species, including industrial strains.
Main Methods:
- Cloning of the ATPsase gene from A. nidulans using complementation of an sC- mutant strain with a gene library.
- Transformation of both A. niger and A. nidulans sC- strains with plasmids containing the cloned ATPsase gene.
Main Results:
- Successful cloning of the ATPsase gene.
- Complementation of sC- mutant strains, restoring their ability to grow on sulfate as the sole sulfur source.
- Demonstration of efficient genetic manipulation strategies in Aspergillus.
Conclusions:
- The cloned ATPsase gene facilitates the genetic engineering of Aspergillus species, including industrially relevant strains.
- The selection system based on ATPsase activity offers a straightforward method for delivering genetic constructs and performing genomic manipulations.