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. 1981 Feb;1(2):136–143. doi: 10.1128/mcb.1.2.136

Electron microscopy of Achlya deoxyribonucleic acid sequence organization.

M Pellegrini 1, W E Timberlake 1, R B Goldberg 1
PMCID: PMC369652  PMID: 6985459

Abstract

Electron microscopic analysis of reassociated deoxyribonucleic acid (DNA) from the aquatic fungus Achlya bisexualis revealed details of the sequence arrangement of the inverted repeats and both the highly and moderately repetitive sequence clusters. We used the gene 32 protein-ethidium bromide technique for visualizing the DNA molecules, a procedure which provides excellent contrast between single- and double-stranded DNA regions. Long (greater than 6-kilobase) DNA fragments were isolated after reannealing to two different repetitive C0t values, and the renatured structures were then visualized in an electron microscope. Our results showed that the inverted repeat sequences were short (0.5 kilobase, number-average) and separated by nonhomologous DNA of various lengths. These pairs of sequences were not clustered within the genome. Both highly repetitive and moderately repetitive DNA sequences were organized as tandem arrays of precisely paired, regularly repeating units. No permuted clusters of repeating sequences were observed, nor was there evidence of interspersion of repetitive with single-copy DNA sequences in the Achlya genome.

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Selected References

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

  1. Bell G. I., DeGennaro L. J., Gelfand D. H., Bishop R. J., Valenzuela P., Rutter W. J. Ribosomal RNA genes of Saccharomyces cerevisiae. I. Physical map of the repeating unit and location of the regions coding for 5 S, 5.8 S, 18 S, and 25 S ribosomal RNAs. J Biol Chem. 1977 Nov 25;252(22):8118–8125. [PubMed] [Google Scholar]
  2. Chamberlin M. E., Britten R. J., Davidson E. H. Sequence organization in Xenopus DNA studied by the electron microscope. J Mol Biol. 1975 Aug 5;96(2):317–333. doi: 10.1016/0022-2836(75)90351-4. [DOI] [PubMed] [Google Scholar]
  3. Chaudhari N., Craig S. P. Internal organization of long repetitive DNA sequences in sea urchin genomes. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6101–6105. doi: 10.1073/pnas.76.12.6101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dons J. J., Wessels J. G. Sequence organization of the nuclear DNA of Schizophyllum commune. Biochim Biophys Acta. 1980 May 30;607(3):385–396. doi: 10.1016/0005-2787(80)90149-5. [DOI] [PubMed] [Google Scholar]
  5. Eslava A. P., Alvarez M. I., Burke P. V., Delbrück M. Genetic recombination in sexual crosses of phycomyces. Genetics. 1975 Jul;80(3):445–462. doi: 10.1093/genetics/80.3.445. [DOI] [PMC free article] [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. Free S. J., Rice P. W., Metzenberg R. L. Arrangement of the genes coding for ribosomal ribonucleic acids in Neurospora crassa. J Bacteriol. 1979 Mar;137(3):1219–1226. doi: 10.1128/jb.137.3.1219-1226.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hereford L. M., Rosbash M. Number and distribution of polyadenylated RNA sequences in yeast. Cell. 1977 Mar;10(3):453–462. doi: 10.1016/0092-8674(77)90032-0. [DOI] [PubMed] [Google Scholar]
  9. Hudspeth M. E., Timberlake W. E., Goldberg R. B. DNA sequence organization in the water mold Achlya. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4332–4336. doi: 10.1073/pnas.74.10.4332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jaworski A. J., Horgen P. A. The ribosomal cistrons of the water mold Achlya bisexualis. Arch Biochem Biophys. 1973 Jul;157(1):260–267. doi: 10.1016/0003-9861(73)90412-8. [DOI] [PubMed] [Google Scholar]
  11. Klein W. H., Thomas T. L., Lai C., Scheller R. H., Britten R. J., Davidson E. H. Characteristics of individual repetitive sequence families in the sea urchin genome studied with cloned repeats. Cell. 1978 Aug;14(4):889–900. doi: 10.1016/0092-8674(78)90344-6. [DOI] [PubMed] [Google Scholar]
  12. Krumlauf R., Marzluf G. A. Characterization of the sequence complexity and organization of the Neurospora crassa genome. Biochemistry. 1979 Aug 21;18(17):3705–3713. doi: 10.1021/bi00584a011. [DOI] [PubMed] [Google Scholar]
  13. Lee C. S., Davis R. W., Davidson N. A physical study by electron microscopy of the terminally reptitious, circularly permuted DNA from the coliphage particles of Escherichia coli 15. J Mol Biol. 1970 Feb 28;48(1):1–22. doi: 10.1016/0022-2836(70)90215-9. [DOI] [PubMed] [Google Scholar]
  14. Rozek C. E., Timberlake W. E. Restriction endonuclease mapping by crossed contact hybridization: the ribosomal RNA genes of Achlya ambisexualis. Nucleic Acids Res. 1979 Nov 24;7(6):1567–1578. doi: 10.1093/nar/7.6.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schmid C. W., Manning J. E., Davidson N. Inverted repeat sequences in the Drosophila genome. Cell. 1975 Jun;5(2):159–172. doi: 10.1016/0092-8674(75)90024-0. [DOI] [PubMed] [Google Scholar]
  16. Smith M. J., Britten R. J., Davidson E. H. Studies on nucleic acid reassociation kinetics: reactivity of single-stranded tails in DNA-DNA renaturation. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4805–4809. doi: 10.1073/pnas.72.12.4805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Timberlake W. E. Low repetitive DNA content in Aspergillus nidulans. Science. 1978 Dec 1;202(4371):973–975. doi: 10.1126/science.362530. [DOI] [PubMed] [Google Scholar]
  18. Timberlake W. E., Shumard D. S., Goldberg R. B. Relationship between nuclear and polysomal RNA populations of Achlya: a simple eucaryotic system. Cell. 1977 Apr;10(4):623–632. doi: 10.1016/0092-8674(77)90095-2. [DOI] [PubMed] [Google Scholar]
  19. Wensink P. C., Tabata S., Pachl C. The clustered and scrambled arrangement of moderately repetitive elements in Drosophila DNA. Cell. 1979 Dec;18(4):1231–1246. doi: 10.1016/0092-8674(79)90235-6. [DOI] [PubMed] [Google Scholar]
  20. Wu M., Davidson N. Use of gene 32 protein staining of single-strand polynucleotides for gene mapping by electron microscopy: application to the phi80d3ilvsu+7 system. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4506–4510. doi: 10.1073/pnas.72.11.4506. [DOI] [PMC free article] [PubMed] [Google Scholar]

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