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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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Abstract.

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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Acknowledgements.

This work was supported by United States Public Health Service Grant GM58770 to R.A.

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Authors and Affiliations

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

    Dong W. Lee & Rodolfo Aramayo

  2. Department of Biology, Graduate School of Arts and Sciences, Washington University, 1 Brookings Drive, Campus Box 1137, St. Louis, MO 63130, USA

    Jeremy R. Haag

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  1. Dong W. Lee
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  2. Jeremy R. Haag
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  3. Rodolfo Aramayo
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Correspondence to Rodolfo Aramayo.

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Communicated by U. Kück

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Lee, D.W., Haag, J.R. & Aramayo, R. Construction of strains for rapid homokaryon purification after integration of constructs at the histidine-3 (his-3) locus of Neurospora crassa . Curr Genet 43, 17–23 (2003). https://doi.org/10.1007/s00294-002-0366-z

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  • DOI: https://doi.org/10.1007/s00294-002-0366-z

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