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Title:   Author  item
                                            SZEWCZYK, EDYTA - University Of Californiaitem
                                            Kasuga, Takaoitem
                                            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/S0167701212004009Citation: 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.


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