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Genetic transformation of Aureobasidium pullulans
1Institute for Microbial and Biochemical Technology, Forest Products Laboratory, U.S. Department of Agriculture, Madison, Wisconsin 53705-2398.
Journal of Biotechnology
|December 1, 1991
Summary
Aureobasidium pullulans was genetically modified using a plasmid conferring hygromycin resistance. The Aspergillus niger glucoamylase promoter showed similar function in both species, despite differing induction by starch.
Area of Science:
- Microbiology
- Molecular Biology
- Genetic Engineering
Background:
- Aureobasidium pullulans is a fungus with industrial applications.
- Genetic transformation is crucial for understanding and manipulating fungal gene expression.
- The hygromycin B phosphotransferase gene (hph) is a common selectable marker.
Purpose of the Study:
- To genetically transform Aureobasidium pullulans strain Y117.
- To investigate the function of the Aspergillus niger glucoamylase (glaA) promoter in A. pullulans.
- To analyze the integration and expression of a foreign gene in A. pullulans.
Main Methods:
- Transformation of A. pullulans with plasmid pDH33 containing the hph gene.
- Southern hybridization to confirm vector integration.
- Analysis of transcriptional start points and promoter activity.
Main Results:
- Successful transformation of A. pullulans with hygromycin resistance.
- Multiple integrated copies of the plasmid were detected in transformants.
- The glaA promoter was functional in A. pullulans but not induced by starch, unlike in A. niger.
- Transcriptional start points were conserved between A. niger and A. pullulans.
Conclusions:
- Aureobasidium pullulans can be genetically transformed using the hph gene and pDH33 plasmid.
- The glaA promoter from Aspergillus niger is active in A. pullulans, with conserved transcriptional start sites.
- Starch does not induce the glaA promoter in A. pullulans, indicating species-specific regulation.