Efficient sequential repetitive gene deletions in Neurospora crassa employing a self-excising β-recombinase/six cassette.

Edyta Szewczyk, Takao Kasuga, Zhiliang Fan

Department of Biological and Agricultural Engineering, University of California, Davis, Davis, CA 95616, USA.

Journal of microbiological methods 2013 Mar

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Despite its long-standing history as a model organism, Neurospora crassa has limited tools for repetitive gene deletions utilizing recyclable self-excising marker systems. Here we describe, for the first time, the functionality of a bacterial recombination system employing β-recombinase acting on six recognition sequences (β-rec/six) in N. crassa, which allowed repetitive site-specific gene deletion and marker recycling. We report generating the mus-51 deletion strain using this system, recycling the marker cassette, and subsequently deleting the global transcriptional regulator gene cre-1. Copyright © 2012 Elsevier B.V. All rights reserved.

Citation

Edyta Szewczyk, Takao Kasuga, Zhiliang Fan. Efficient sequential repetitive gene deletions in Neurospora crassa employing a self-excising β-recombinase/six cassette. Journal of microbiological methods. 2013 Mar;92(3):236-43