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Enzyme and Microbial Technology
Vol. 51, No. 1, 2012; Pages: 53–58


A novel selectable marker based on Aspergillus niger arginase expression

Kashyap Dave, Manmeet Ahuja, T.N. Jayashri, Rekha Bisht Sirola, Narayan S. Punekar

Biotechnology Group, Department of Bioscience and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai 400 076, India.

Abstract

Selectable markers are valuable tools in transforming asexual fungi like Aspergillus niger. An arginase (agaA) expression vector and a suitable arginase-disrupted host would define a novel nutritional marker/selection for transformation. The development of such a marker was successfully achieved in two steps. The single genomic copy of A. niger arginase gene was disrupted by homologous integration of the bar marker. The agaA disruptant was subsequently complemented by transforming it with agaA expression vectors. Both citA and trpC promoters were able to drive the expression of arginase cDNA. Such agaA+ transformants displayed arginase expression pattern distinct from that of the parent strain. The results are also consistent with a single catabolic route for arginine in this fungus. A simple yet novel arginine-based selection for filamentous fungal transformation is thus described.

Keywords:Fungal transformation,Arginase (agaA) disruption,Nutritional markers,Aspergillus niger.


 
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