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. 1995 Apr;139(4):1545–1551. doi: 10.1093/genetics/139.4.1545

Some Property of the Nucleus Determines the Competence of Neurospora Crassa for Transformation

J Grotelueschen 1, R L Metzenberg 1
PMCID: PMC1206483  PMID: 7789759

Abstract

In Neurospora, transformation of spheroplasts is quite efficient and usually occurs with the transforming DNA integrated at ectopic sites in the chromosome. However, only a small fraction of the spheroplasts is actually competent for transformation. To distinguish whether the limitation to competence is at the level of the plasma membrane or at the level of the nucleus, we performed experiments in which heterocaryotic spheroplasts were required to integrate two different plasmids in one transformation procedure. The cotransformants were then analyzed to determine into which nucleus or nuclei the separate plasmids had integrated. Results of such experiments confirm that successful ectopic transformation in Neurospora crassa requires a competent nucleus. The integration patterns of the two separate plasmids indicate that the availability of appropriate chromosomal sites for ectopic integration may be an aspect of nuclear competence.

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Selected References

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  1. Akins R. A., Lambowitz A. M. General method for cloning Neurospora crassa nuclear genes by complementation of mutants. Mol Cell Biol. 1985 Sep;5(9):2272–2278. doi: 10.1128/mcb.5.9.2272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Asch D. K., Frederick G., Kinsey J. A., Perkins D. D. Analysis of junction sequences resulting from integration at nonhomologous loci in Neurospora crassa. Genetics. 1992 Apr;130(4):737–748. doi: 10.1093/genetics/130.4.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barry E. G., Perkins D. D. Position of linkage group V markers in chromosome 2 of Neurospora crassa. J Hered. 1969 May-Jun;60(3):120–125. doi: 10.1093/oxfordjournals.jhered.a107952. [DOI] [PubMed] [Google Scholar]
  4. Butler D. K., Metzenberg R. L. Premeiotic change of nucleolus organizer size in Neurospora. Genetics. 1989 Aug;122(4):783–791. doi: 10.1093/genetics/122.4.783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Butler D. K. Ribosomal DNA is a site of chromosome breakage in aneuploid strains of Neurospora. Genetics. 1992 Jul;131(3):581–592. doi: 10.1093/genetics/131.3.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chu G., Vollrath D., Davis R. W. Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science. 1986 Dec 19;234(4783):1582–1585. doi: 10.1126/science.3538420. [DOI] [PubMed] [Google Scholar]
  7. Pandit N. N., Russo V. E. Reversible inactivation of a foreign gene, hph, during the asexual cycle in Neurospora crassa transformants. Mol Gen Genet. 1992 Sep;234(3):412–422. doi: 10.1007/BF00538700. [DOI] [PubMed] [Google Scholar]
  8. Perkins D. D., Barry E. G. The cytogenetics of Neurospora. Adv Genet. 1977;19:133–285. doi: 10.1016/s0065-2660(08)60246-1. [DOI] [PubMed] [Google Scholar]
  9. Perkins D. D., Kinsey J. A., Asch D. K., Frederick G. D. Chromosome rearrangements recovered following transformation of Neurospora crassa. Genetics. 1993 Jul;134(3):729–736. doi: 10.1093/genetics/134.3.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Perkins D. D., Radford A., Newmeyer D., Björkman M. Chromosomal loci of Neurospora crassa. Microbiol Rev. 1982 Dec;46(4):426–570. doi: 10.1128/mr.46.4.426-570.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Perkins D. D. The manifestation of chromosome rearrangements in unordered asci of Neurospora. Genetics. 1974 Jul;77(3):459–489. doi: 10.1093/genetics/77.3.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Reid L. H., Shesely E. G., Kim H. S., Smithies O. Cotransformation and gene targeting in mouse embryonic stem cells. Mol Cell Biol. 1991 May;11(5):2769–2777. doi: 10.1128/mcb.11.5.2769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Robins D. M., Ripley S., Henderson A. S., Axel R. Transforming DNA integrates into the host chromosome. Cell. 1981 Jan;23(1):29–39. doi: 10.1016/0092-8674(81)90267-1. [DOI] [PubMed] [Google Scholar]
  14. Schiestl R. H., Dominska M., Petes T. D. Transformation of Saccharomyces cerevisiae with nonhomologous DNA: illegitimate integration of transforming DNA into yeast chromosomes and in vivo ligation of transforming DNA to mitochondrial DNA sequences. Mol Cell Biol. 1993 May;13(5):2697–2705. doi: 10.1128/mcb.13.5.2697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Serna L., Stadler D. Nuclear division cycle in germinating conidia of Neurospora crassa. J Bacteriol. 1978 Oct;136(1):341–351. doi: 10.1128/jb.136.1.341-351.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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