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Efficient Transfection of Neurospora Crassa

Katherine A. Borkovich PhD and Kimberly Vollmer
Katherine A. Borkovich PhD, University of Texas-Houston Medical Center,
Dept. of Microbiology and Molecular Genetics, Houston, Texas.
Kimberly Vollmer, Brinkmann™ Instruments Inc., BioSystems Application Lab, Westbury, New York. INTRODUCTION:

The diverse reproductive capabilities and the nutritional conventions of Neurospora crassa have made this filamentous fungus a model organism in the field of molecular biology and genetics for more than a half a century. In fact, N. crassa was used in the Nobel Prize-winning "One-gene One-enzyme" theory [1]. Since then, N. crassa has been used as a model organism in other types of molecular and genetic applications, joining the illustrious company of E. coli and Drosphila melanogaster.

Perhaps one of the most important roles forN. crassa has been as a host organism for exogenous DNA. Over the years, several techniques have been developed to introduce exogenous material into cells. Electroporation, a technique utilizing electronically regulated pulses to open the cellular membrane and introduce material, has become an easy and efficient method of transfection.

The Eppendorf® Electroporator 2510 employs electric pulse technology to provide an easy and efficient means of introducing exogenous DNA into N. crassa conidia [2]. Most procedures in the past have either used protoplasts for the transfection of N. crassa, or have required the addition of an enzyme to facilitate the weakening of the cell wall. When using the Electroporator 2510, germinated conidia can
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