|SZEWCZYK, EDYTA - University Of California|
|FAN, ZHILIANG - University Of California|
Submitted to: Journal of Microbiological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/10/2013
Publication Date: 3/1/2013
Publication URL: http://www.sciencedirect.com/science/article/pii/S0167701212004009
Citation: Szewczyk, E., Kasuga, T., Fan, Z. 2013. Efficient sequential repetitive gene deletions in Neurospora crassa. Journal of Microbiological Methods. 92:236-243.
Interpretive Summary: A plant decomposing fungus, Neurospora crassa, has been used for the study of biochemistry and cellular biology. In this study, we have developed a molecular biological method, which allows us to sequentially manipulate genes to create desirable genotypes. This method is highly valuable for bioengineering as well as functional genomics study.
Technical Abstract: 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 ß-recombinase acting on six recognition sequences (ß-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.