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. 1990 Nov;87(21):8247–8251. doi: 10.1073/pnas.87.21.8247

Electrophoretic karyotype for Dictyostelium discoideum.

E C Cox 1, C D Vocke 1, S Walter 1, K Y Gregg 1, E S Bain 1
PMCID: PMC54932  PMID: 2236037

Abstract

This paper reports on the separation of the Dictyostelium discoideum chromosomes by pulse-field electrophoresis and the correlation of the electrophoretic pattern with linkage groups established by classical genetic methods. In two commonly used laboratory strains, five chromosome-sized DNA molecules have been identified. Although the majority of the molecular probes used in this study can be unambiguously assigned to established linkage groups, the electrophoretic karyotype differs between the closely related strains AX3k and NC4, suggesting that chromosomal fragmentation may have occurred during their maintenance and growth. The largest chromosome identified in this study is approximately 9 million base pairs. To achieve resolution with molecules of this size, programmed voltage gradients were used in addition to programmed pulse times.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. André E., Brink M., Gerisch G., Isenberg G., Noegel A., Schleicher M., Segall J. E., Wallraff E. A Dictyostelium mutant deficient in severin, an F-actin fragmenting protein, shows normal motility and chemotaxis. J Cell Biol. 1989 Mar;108(3):985–995. doi: 10.1083/jcb.108.3.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. André E., Lottspeich F., Schleicher M., Noegel A. Severin, gelsolin, and villin share a homologous sequence in regions presumed to contain F-actin severing domains. J Biol Chem. 1988 Jan 15;263(2):722–727. [PubMed] [Google Scholar]
  3. Ashworth J. M., Watts D. J. Metabolism of the cellular slime mould Dictyostelium discoideum grown in axenic culture. Biochem J. 1970 Sep;119(2):175–182. doi: 10.1042/bj1190175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carle G. F., Olson M. V. Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis. Nucleic Acids Res. 1984 Jul 25;12(14):5647–5664. doi: 10.1093/nar/12.14.5647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cockburn A. F., Taylor W. C., Firtel R. A. Dictyostelium rDNA consists of non-chromosomal palindromic dimers containing 5S and 36S coding regions. Chromosoma. 1978 Dec 21;70(1):19–29. doi: 10.1007/BF00292212. [DOI] [PubMed] [Google Scholar]
  8. Cohen S. M., Knecht D., Lodish H. F., Loomis W. F. DNA sequences required for expression of a Dictyostelium actin gene. EMBO J. 1986 Dec 1;5(12):3361–3366. doi: 10.1002/j.1460-2075.1986.tb04651.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cole R. A., Williams K. L. Insertion of transformation vector DNA into different chromosomal sites of Dictyostelium discoideum as determined by pulse field electrophoresis. Nucleic Acids Res. 1988 Jun 10;16(11):4891–4902. doi: 10.1093/nar/16.11.4891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. De Lozanne A., Spudich J. A. Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination. Science. 1987 May 29;236(4805):1086–1091. doi: 10.1126/science.3576222. [DOI] [PubMed] [Google Scholar]
  11. DeLozanne A., Lewis M., Spudich J. A., Leinwand L. A. Cloning and characterization of a nonmuscle myosin heavy chain cDNA. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6807–6810. doi: 10.1073/pnas.82.20.6807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Devreotes P. Dictyostelium discoideum: a model system for cell-cell interactions in development. Science. 1989 Sep 8;245(4922):1054–1058. doi: 10.1126/science.2672337. [DOI] [PubMed] [Google Scholar]
  13. Dingermann T., Amon-Böhm E., Bertling W., Marschalek R., Nerke K. A family of non-allelic tRNA(ValGUU) genes from the cellular slime mold Dictyostelium discoideum. Gene. 1988 Dec 20;73(2):373–384. doi: 10.1016/0378-1119(88)90502-1. [DOI] [PubMed] [Google Scholar]
  14. Dingermann T., Amon E., Williams K. L., Welker D. L. Chromosomal mapping of tRNA genes from Dictyostelium discoideum. Mol Gen Genet. 1987 Apr;207(1):176–187. doi: 10.1007/BF00331507. [DOI] [PubMed] [Google Scholar]
  15. Dingermann T., Bertling W., Pistel F., Amon E. Characterisation of a Dictyostelium discoideum DNA fragment coding for a putative tRNAValGUU gene. Evidence for a single transcription unit consisting of two overlapping class III genes. Eur J Biochem. 1985 Jan 15;146(2):449–458. doi: 10.1111/j.1432-1033.1985.tb08672.x. [DOI] [PubMed] [Google Scholar]
  16. Early A. E., Williams J. G., Meyer H. E., Por S. B., Smith E., Williams K. L., Gooley A. A. Structural characterization of Dictyostelium discoideum prespore-specific gene D19 and of its product, cell surface glycoprotein PsA. Mol Cell Biol. 1988 Aug;8(8):3458–3466. doi: 10.1128/mcb.8.8.3458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Grant W. N., Welker D. L., Williams K. L. A polymorphic, prespore-specific cell surface glycoprotein is present in the extracellular matrix of Dictyostelium discoideum. Mol Cell Biol. 1985 Oct;5(10):2559–2566. doi: 10.1128/mcb.5.10.2559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Klein P. S., Sun T. J., Saxe C. L., 3rd, Kimmel A. R., Johnson R. L., Devreotes P. N. A chemoattractant receptor controls development in Dictyostelium discoideum. Science. 1988 Sep 16;241(4872):1467–1472. doi: 10.1126/science.3047871. [DOI] [PubMed] [Google Scholar]
  19. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  20. Levene S. D., Zimm B. H. Separations of open-circular DNA using pulsed-field electrophoresis. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4054–4057. doi: 10.1073/pnas.84.12.4054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maizels N. Dictyostelium 17S, 25S, and 5S rDNAs lie within a 38,000 base pair repeated unit. Cell. 1976 Nov;9(3):431–438. doi: 10.1016/0092-8674(76)90088-x. [DOI] [PubMed] [Google Scholar]
  22. McDonough J. P., Springer W. R., Barondes S. H. Species-specific cell cohesion in cellular slime molds. Demonstration by several quantitative assays and with multiple species. Exp Cell Res. 1980 Jan;125(1):1–14. doi: 10.1016/0014-4827(80)90182-2. [DOI] [PubMed] [Google Scholar]
  23. Ness P. J., Labhart P., Banz E., Koller T., Parish R. W. Chromatin structure along the ribosomal DNA of Dictyostelium. Regional differences and changes accompanying cell differentiation. J Mol Biol. 1983 May 25;166(3):361–381. doi: 10.1016/s0022-2836(83)80090-4. [DOI] [PubMed] [Google Scholar]
  24. Noegel A., Gerisch G., Stadler J., Westphal M. Complete sequence and transcript regulation of a cell adhesion protein from aggregating Dictyostelium cells. EMBO J. 1986 Jul;5(7):1473–1476. doi: 10.1002/j.1460-2075.1986.tb04384.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pears C. J., Mahbubani H. M., Williams J. G. Characterization of two highly diverged but developmentally co-regulated cysteine proteinase genes in Dictyostelium discoideum. Nucleic Acids Res. 1985 Dec 20;13(24):8853–8866. doi: 10.1093/nar/13.24.8853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Podgorski G. J., Faure M., Franke J., Kessin R. H. The cyclic nucleotide phosphodiesterase of Dictyostelium discoideum: the structure of the gene and its regulation and role in development. Dev Genet. 1988;9(4-5):267–278. doi: 10.1002/dvg.1020090409. [DOI] [PubMed] [Google Scholar]
  27. Pupillo M., Klein P., Vaughan R., Pitt G., Lilly P., Sun T., Devreotes P., Kumagai A., Firtel R. cAMP receptor and G-protein interactions control development in Dictyostelium. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 2):657–665. doi: 10.1101/sqb.1988.053.01.075. [DOI] [PubMed] [Google Scholar]
  28. Schwartz D. C., Saffran W., Welsh J., Haas R., Goldenberg M., Cantor C. R. New techniques for purifying large DNAs and studying their properties and packaging. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 1):189–195. doi: 10.1101/sqb.1983.047.01.024. [DOI] [PubMed] [Google Scholar]
  29. Smith C. L., Matsumoto T., Niwa O., Klco S., Fan J. B., Yanagida M., Cantor C. R. An electrophoretic karyotype for Schizosaccharomyces pombe by pulsed field gel electrophoresis. Nucleic Acids Res. 1987 Jun 11;15(11):4481–4489. doi: 10.1093/nar/15.11.4481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tsang A. S., Devine J. M., Williams J. G. The multiple subunits of discoidin I are encoded by different genes. Dev Biol. 1981 May;84(1):212–217. doi: 10.1016/0012-1606(81)90385-7. [DOI] [PubMed] [Google Scholar]
  31. Wallace J. S., Newell P. C. Genetic analysis by mitotic recombination in Dictyostelium discoideum of growth and developmental loci on linkage group VII. J Gen Microbiol. 1982 May;128(5):953–964. doi: 10.1099/00221287-128-5-953. [DOI] [PubMed] [Google Scholar]
  32. Wallraff E., Schleicher M., Modersitzki M., Rieger D., Isenberg G., Gerisch G. Selection of Dictyostelium mutants defective in cytoskeletal proteins: use of an antibody that binds to the ends of alpha-actinin rods. EMBO J. 1986 Jan;5(1):61–67. doi: 10.1002/j.1460-2075.1986.tb04178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Warren A. J., Warren W. D., Cox E. C. Genetic and morphological study of aggregation in the cellular slime mold Polysphondylium violaceum. Genetics. 1976 May;83(1):25–47. doi: 10.1093/genetics/83.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Welker D. L., De Lozanne A., Spudich J. A. Linkage analysis of the myosin heavy chain gene in Dictyostelium discoideum using a mutation generated by homologous recombination. Mol Gen Genet. 1989 Apr;216(2-3):498–502. doi: 10.1007/BF00334396. [DOI] [PubMed] [Google Scholar]
  35. Welker D. L., Hirth K. P., Romans P., Noegel A., Firtel R. A., Williams K. L. The use of restriction fragment length polymorphisms and DNA duplications to study the organization of the actin multigene family in Dictyostelium discoideum. Genetics. 1986 Jan;112(1):27–42. doi: 10.1093/genetics/112.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Welker D. L. The discoidin I gene family of Dictyostelium discoideum is linked to genes regulating its expression. Genetics. 1988 Jul;119(3):571–578. doi: 10.1093/genetics/119.3.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Welker D. L., Williams K. L. A genetic map of Dictyostelium discoideum based on mitotic recombination. Genetics. 1982 Dec;102(4):691–710. doi: 10.1093/genetics/102.4.691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Williams J. G., North M. J., Mahbubani H. A developmentally regulated cysteine proteinase in Dictyostelium discoideum. EMBO J. 1985 Apr;4(4):999–1006. doi: 10.1002/j.1460-2075.1985.tb03730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Williams J. G. The role of diffusible molecules in regulating the cellular differentiation of Dictyostelium discoideum. Development. 1988 May;103(1):1–16. [PubMed] [Google Scholar]

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