Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Dec;79(23):7356–7360. doi: 10.1073/pnas.79.23.7356

A DNA-mediated transformation system for Dictyostelium discoideum.

K P Hirth, C A Edwards, R A Firtel
PMCID: PMC347338  PMID: 6296830

Abstract

We have established a transformation system for Dictyostelium discoideum. The transformation vector contains the protein coding region of the Tn5 neomycin resistance gene fused to the proposed promoter of the Dictyostelium actin 8 gene; the vector also contains a sequence that acts as an autonomously replicating sequence (ars) in yeast. Using this vector, we can transform Dictyostelium vegetative amoebae to be resistant to aminoglycoside G418 at a frequency of between 10(-6) and 10(-4) of the input cells. The transformed cell lines are stable and contain vector sequences integrated within chromosomal DNA.

Full text

PDF
7356

Images in this article

7358Image
on p.7358
7359Image
on p.7359
7359Image
on p.7359

Selected References

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

  1. Chung S., Landfear S. M., Blumberg D. D., Cohen N. S., Lodish H. F. Synthesis and stability of developmentally regulated dictyostelium mRNAs are affected by cell--cell contact and cAMP. Cell. 1981 Jun;24(3):785–797. doi: 10.1016/0092-8674(81)90104-5. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Cocucci S. M., Sussman M. RNA in cytoplasmic and nuclear fractions of cellular slime mold amebas. J Cell Biol. 1970 May;45(2):399–407. doi: 10.1083/jcb.45.2.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Colbère-Garapin F., Horodniceanu F., Kourilsky P., Garapin A. C. A new dominant hybrid selective marker for higher eukaryotic cells. J Mol Biol. 1981 Jul 25;150(1):1–14. doi: 10.1016/0022-2836(81)90321-1. [DOI] [PubMed] [Google Scholar]
  5. Firtel R. A., Bonner J. Characterization of the genome of the cellular slime mold Dictyostelium discoideum. J Mol Biol. 1972 May 28;66(3):339–361. doi: 10.1016/0022-2836(72)90419-6. [DOI] [PubMed] [Google Scholar]
  6. Firtel R. A., Cockburn A., Frankel G., Hershfield V. Structural organization of the genome of Dictyostelium discoideum: analysis by EcoR1 restriction endonuclease. J Mol Biol. 1976 Apr 25;102(4):831–852. doi: 10.1016/0022-2836(76)90294-1. [DOI] [PubMed] [Google Scholar]
  7. Gross J. D., Town C. D., Brookman J. J., Jermyn K. A., Peacey M. J., Kay R. R. Cell patterning in Dictyostelium. Philos Trans R Soc Lond B Biol Sci. 1981 Oct 7;295(1078):497–508. doi: 10.1098/rstb.1981.0156. [DOI] [PubMed] [Google Scholar]
  8. Hinnen A., Hicks J. B., Fink G. R. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. doi: 10.1073/pnas.75.4.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jimenez A., Davies J. Expression of a transposable antibiotic resistance element in Saccharomyces. Nature. 1980 Oct 30;287(5785):869–871. doi: 10.1038/287869a0. [DOI] [PubMed] [Google Scholar]
  10. Kindle K. L., Firtel R. A. Identification and analysis of Dictyostelium actin genes, a family of moderately repeated genes. Cell. 1978 Nov;15(3):763–778. doi: 10.1016/0092-8674(78)90262-3. [DOI] [PubMed] [Google Scholar]
  11. Kingsman A. J., Clarke L., Mortimer R. K., Carbon J. Replication in Saccharomyces cerevisiae of plasmid pBR313 carrying DNA from the yeast trpl region. Gene. 1979 Oct;7(2):141–152. doi: 10.1016/0378-1119(79)90029-5. [DOI] [PubMed] [Google Scholar]
  12. Kiss G. B., Amin A. A., Pearlman R. E. Two separate regions of the extrachromosomal ribosomal deoxyribonucleic acid of Tetrahymena thermophila enable autonomous replication of plasmids in Saccharomyces cerevisiae. Mol Cell Biol. 1981 Jun;1(6):535–543. doi: 10.1128/mcb.1.6.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Loomis W. F., Jr Sensitivity of Dictyostelium discoideum to nucleic acid analogues. Exp Cell Res. 1971 Feb;64(2):484–486. doi: 10.1016/0014-4827(71)90107-8. [DOI] [PubMed] [Google Scholar]
  14. Lusky M., Botchan M. Inhibition of SV40 replication in simian cells by specific pBR322 DNA sequences. Nature. 1981 Sep 3;293(5827):79–81. doi: 10.1038/293079a0. [DOI] [PubMed] [Google Scholar]
  15. McKeown M., Firtel R. A. Differential expression and 5' end mapping of actin genes in Dictyostelium. Cell. 1981 Jun;24(3):799–807. doi: 10.1016/0092-8674(81)90105-7. [DOI] [PubMed] [Google Scholar]
  16. McKeown M., Firtel R. A. Evidence for sub-families of actin genes in Dictyostelium as determined by comparisons of 3' end sequences. J Mol Biol. 1981 Oct 5;151(4):593–606. doi: 10.1016/0022-2836(81)90425-3. [DOI] [PubMed] [Google Scholar]
  17. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. doi: 10.1073/pnas.78.10.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Poole S., Firtel R. A., Lamar E., Rowekamp W. Sequence and expression of the discoidin I gene family in Dictyostelium discoideum. J Mol Biol. 1981 Dec 5;153(2):273–289. doi: 10.1016/0022-2836(81)90278-3. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Rowekamp W., Poole S., Firtel R. A. Analysis of the multigene family coding the developmentally regulated carbohydrate-binding protein discoidin-I in D. discoideum. Cell. 1980 Jun;20(2):495–505. doi: 10.1016/0092-8674(80)90636-4. [DOI] [PubMed] [Google Scholar]
  21. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  22. Stinchcomb D. T., Thomas M., Kelly J., Selker E., Davis R. W. Eukaryotic DNA segments capable of autonomous replication in yeast. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4559–4563. doi: 10.1073/pnas.77.8.4559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035–1039. doi: 10.1073/pnas.76.3.1035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Williams J. G., Tsang A. S., Mahbubani H. A change in the rate of transcription of a eukaryotic gene in response to cyclic AMP. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7171–7175. doi: 10.1073/pnas.77.12.7171. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES