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Current Genetics

, Volume 29, Issue 3, pp 301–305 | Cite as

Mapping translocation breakpoints by orthogonal field agarose-gel electrophoresis

  • M. L. Smith
  • N. L. Glass
Original Paper

Abstract

Orthogonal field agarose-gel electrophoresis (OFAGE) of chromosomes from translocation-bearing and normalNeurospora crassa strains was utilized, first, to recover cosmids from a translocated region, and second, to map translocation breakpoints. Surprisingly, the right breakpoints in two independently derived, interstitial translocations,T(II → III) AR18 andT(II → VI)P2869, are within about 5.6 kbp of each other suggesting that this region of linkage group (LG) II may be fragile or otherwise subject to chromosome breakage. Mapping translocation breakpoints through OFAGE, or other similar methods, should allow for DNA sequencing across breakpoints that are not associated with mutant phenotypes or that are not within walking distance of cloned markers.

Key words

Translocation breakpoint Neurospora crassa OFAGE Electrophoretic karyotype 

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References

  1. Antonarakis SE (1993) Human chromosome 21: genome mapping and exploration, circa 1993. Trends Genet 9:142–148PubMedGoogle Scholar
  2. Chen SJ, Chen Z, Font M-P, d'Auriol L, Larsen C-J, Berger R (1989) Structural alterations of the BCR and ABL genes in Ph1-positive acute leukemias with rearrangements in the BCR gene first intron: further evidence implicating Alu sequences in the chromosome translocation. Nucleic Acids Res 17:7631–7642PubMedGoogle Scholar
  3. Cirera S, Martin-Campos JM, Segarra C, Aguadé M (1995) Molecular characterization of the breakpoints of an inversion fixed betweenDrosophila melanogaster andD. subobscura Genetics 139:321–326PubMedGoogle Scholar
  4. Deininger PI (1989) SINEs: short interspersed repeated elements in higher eukaryotes. In: Berg DE, Howe MM (eds) Mobile DNA. American Society for Microbiology, Washington DCGoogle Scholar
  5. Döring H-P, Nelsen-Salz B, Garber R, Tillmann E (1989) DoubleDs elements are involved in specific chromosome breakage. Mol Gen Genet 219:299–305PubMedGoogle Scholar
  6. Engels WR, Preston CR (1984) Formation of chromosome rearrangements byP factors in Drosophila. Genetics 107:657–678PubMedGoogle Scholar
  7. Glass NL, Smith ML (1994) Structure and function of a mating-type gene from the homothallic speciesNeurospora africana. Mol Gen Genet 244:401–409PubMedGoogle Scholar
  8. Kang S, Metzenberg RL (1990) Molecular analysis ofnuc-1 +, a gene controlling phosphorus acquisition inNeurospora crassa. Mol Cell Biol 10:5839–5848PubMedGoogle Scholar
  9. Kinsey JA, Helber J (1989) Isolation of a transposable element fromNeurospora crassa. Proc Natl Acad Sci USA 86:1929–1933PubMedGoogle Scholar
  10. Krumlauf R, Marzluf GA (1980) Genome organization and characterization of the repetitive and inverted repeat DNA sequences inNeurospora crassa. J Biol Chem 255:1138–1145PubMedGoogle Scholar
  11. Longo E, Vezinhet F (1993) Chromosome rearrangements during vegetative growth of a wild strain ofSaccharomyces cerevisiae. Appl Environ Microbiol 59:322–326PubMedGoogle Scholar
  12. Metzenberg RL, Grotelueschen J (1992) Restriction polymorphism maps ofNeurospora crassa: update. Fungal Genet Newslett 39:50–58Google Scholar
  13. Metzenberg RL, Stevens JN, Selker EU, Morzycka-Wroblewska E (1985) Identification and chromosomal distribution of 5s RNA genes inNeurospora crassa. Proc Natl Acad Sci USA 82:2067–2071PubMedGoogle Scholar
  14. Orbach MJ (1992) Fungal Genet Newslett 39:92Google Scholar
  15. Orbach MJ (1994) A cosmid with a HyR marker for fungal library construction and screening. Gene 150:159–162PubMedGoogle Scholar
  16. Orbach MJ, Vollrath D, Davis RW, Yanofsky C (1988) An electrophoretic karyotype ofNeurospora crassa. Mol Cell Biol 8:1469–1473PubMedGoogle Scholar
  17. Paluh JL, Plamann M, Krüger D, Barthelmess IB, Yanofsky C, Perkins DD (1990) Determination of the inactivating alteration in two mutant alleles of theNeurospora crassa cross-pathway control genecpc-1. Genetics 124:599–606PubMedGoogle Scholar
  18. Perkins DD (1986) Determining the order of genes, centromeres, and rearrangement breakpoints inNeurospora by tests of duplication coverage. J Genet 65:121–144Google Scholar
  19. Perkins DD (1992)Neurospora crassa genetic maps. Fungal Genet Newslett 39:153–162Google Scholar
  20. Perkins DD, Barry EG (1977) The cytogenetics ofNeurospora. Adv Genet 19:133–285PubMedGoogle Scholar
  21. Perkins DD, Radford A, Newmeyer D, Björkman M (1982) Chromosomal loci ofNeurospora crassa. Microbiol Rev 46:426–570PubMedGoogle Scholar
  22. Perkins DD, Kinsey JA, Asch DK, Frederick GD (1993a) Chromosome rearrangements recovered following transformation ofNeurospora crassa. Genetics 134:729–736PubMedGoogle Scholar
  23. Perkins DD, Leslie JF, Jacobson DJ (1993b) Strains for identifying and studying individual vegetative (heterokaryon) incompatibility loci inNeurospora crassa. Fungal Genet Newslett 40:69–73Google Scholar
  24. Sambrook J, Fritsch EF, Maniatis, T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  25. Schmidhauser TJ, Lauter FR, Russo VEA, Yanofsky C (1990) Cloning, sequence and photoregulation ofal-1, a carotenoid biosynthetic gene ofNeurospora crassa. Mol Cell Biol 10:5064–5070PubMedGoogle Scholar
  26. Skinner DZ, Budde AD, Leong SA (1991) Molecular karyotype analysis of fungi. In: Bennett JW, Lasure LL (eds) More gene manipulations in fungi. Academic Press Inc, TorontoGoogle Scholar
  27. Tóth G, Jurka J (1994) Repetitive DNA in and around translocation breakpoints of the Philadelphia chromosome. Gene 140:285–288PubMedGoogle Scholar
  28. Tsubota SI, Rosenberg D, Szostak H, Rubin D, Schedl P (1989) The cloning of theBar region and theB breakpoint inDrosophila melanogaster: evidence for a transposon-induced rearrangement. Genetics 122:881–890PubMedGoogle Scholar
  29. Vogel HJ (1964) Distribution of lysine pathways among fungi: evolutionary implications. Am Nat 98:435–446Google Scholar
  30. Vollmer SJ, Yanofsky C (1986) Efficient cloning of genes inNeurospora crassa. Proc Natl Acad Sci USA 83:4869–4873Google Scholar
  31. Wesley CS, Eanes WF (1994) Isolation and analysis of the breakpoint sequences of chromosome inversionIn(3L)Payne inDrosophila melanogaster. Proc Natl Acad Sci USA 91:3132–3136PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • M. L. Smith
    • 1
  • N. L. Glass
    • 1
  1. 1.Department of Botany and Biotechnology LaboratoryUniversity of British ColumbiaVancouverCanada

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