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A novel selectable marker based on Aspergillus niger arginase expression.

Kashyap Dave¹, Manmeet Ahuja, T N Jayashri

¹Biotechnology Group, Department of Bioscience and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India.

Enzyme and Microbial Technology

|May 15, 2012

Summary

A novel arginine-based selection method was developed for Aspergillus niger transformation. This method uses an arginase-disrupted host and expression vector, enabling efficient genetic modification of asexual fungi.

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Area of Science:

Biotechnology
Molecular Biology
Mycology

Background:

Selectable markers are crucial for genetic transformation in asexual fungi like Aspergillus niger.
Developing novel nutritional markers can enhance transformation efficiency and simplify selection processes.

Purpose of the Study:

To develop and describe a novel arginine-based nutritional marker and selection system for Aspergillus niger transformation.
To create an arginase-disrupted host strain and an arginase (agaA) expression vector for this new selection system.

Main Methods:

Disruption of the single genomic copy of the Aspergillus niger arginase (agaA) gene using homologous integration of the bar marker.
Complementation of the agaA disruptant strain by transforming it with agaA expression vectors driven by citA and trpC promoters.

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Analysis of arginase expression patterns in transformants compared to the parent strain.

Main Results:

Successful disruption of the A. niger arginase gene and creation of a functional arginase-disrupted host.
Demonstration that both citA and trpC promoters can drive arginase cDNA expression, complementing the disrupted gene.
Observation of distinct arginase expression patterns in complemented transformants, supporting a single arginine catabolic route.

Conclusions:

A simple and novel arginine-based selection system for filamentous fungal transformation has been successfully developed.
This method provides a valuable new tool for the genetic manipulation of asexual fungi, specifically Aspergillus niger.
The findings confirm the utility of nutritional markers and provide insights into arginine metabolism in Aspergillus niger.