Cloning and use of sC as homologous marker for Aspergillus niger transformation

The sC sequence from Aspergillus niger was cloned and developed into a homologous marker system for genetic transformation. The coding region of the sC gene amplified by PCR from the A. niger genome was provided with Aspergillus nidulans expression signals ( gpdA promoter and trpC terminator). This...

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Bibliographic Details
Published in:Journal of microbiological methods 2005-05, Vol.61 (2), p.219-224
Main Authors: Varadarajalu, Lakshmi P., Punekar, Narayan S.
Format: Article
Language:English
Subjects:
Aspergillus nidulans
Aspergillus nidulans - genetics
Aspergillus niger
Aspergillus niger - enzymology
Aspergillus niger - genetics
ATP sulfurylase activity
Biological and medical sciences
Blotting, Southern
Cloning, Molecular
DNA, Fungal - genetics
Fundamental and applied biological sciences. Psychology
Genetic Markers - genetics
Genetic Markers - physiology
Marker
Microbiology
Miscellaneous
Mycology
Polymerase Chain Reaction
sC gene
Sulfate Adenylyltransferase - genetics
Sulfate Adenylyltransferase - metabolism
Transformation
Transformation, Genetic - physiology
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Summary:The sC sequence from Aspergillus niger was cloned and developed into a homologous marker system for genetic transformation. The coding region of the sC gene amplified by PCR from the A. niger genome was provided with Aspergillus nidulans expression signals ( gpdA promoter and trpC terminator). This chimeric construct was used to successfully transform a spontaneous sC − isolate of A. niger to prototrophy. The transformants analyzed by Southern analysis showed integration of multiple copies of the transforming DNA. They also exhibited much higher ATP sulfurylase activity than the wild-type A. niger strain reinforcing the molecular data. This demonstrates the usefulness of the sC niger construct, driven by PgpdA, as a marker for A. niger transformation.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2004.11.022