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Curr Genet. 2003 Apr;43(1):17-23. Epub 2003 Feb 18.

Construction of strains for rapid homokaryon purification after integration of constructs at the histidine-3 ( his-3) locus of Neurospora crassa.

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Department of Biology, College of Science, Texas A&M University, Room 415, Building BSBW, College Station, TX 77843-3258, USA.


We report the construction of histidine-3 (his-3) strains of Neurospora crassa containing the hygromycin B phosphotransferase gene of Escherichia coli (hph(+)) fused in-frame to the herpes simplex virus thymidine kinase gene (tk(+); Lupton et al. 1991), integrated at the his-3 locus. We also report the construction of two ampicillin-resistant and two kanamycin-resistant his-3 gene-replacement vector plasmids. The combined use of these strains and plasmids for his-3-targeted gene integration allows for the rapid identification of homokaryotic transformants containing the expected gene replacement event.

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