Isolation and Analysis of Genomic Sequences from Mycorrhizal Fungi

  • M. G. De Santo
  • S. Filosa
  • A. Franzè
  • G. Martini

Abstract

The isolation of effective probes is of paramount importance in studies aimed at elucidating the molecular mechanisms underlying the development of ectomycorrhizal fungi, as well as in the creation of biotechnological tools for improving truffle harvest and species identification. In one strategy which we are using to isolate DNA probes, recombinant plasmids are randomly isolated from genomic libraries and their nucleotide sequences compared with available data banks of nucleotide and aminoacid sequences. As a first step in this strategy, we have analyzed 56 clones from a genomic library of plasmids bearing small inserts of DNA from Tuber albidum. Several clones were singled out on the basis of sequence similarity with known genes. Among these was a recombinant plasmid carrying nucleotide sequences potentially coding for a protein extremely similar to the a subunit of Fatty Acid Synthase of other fungal species. The potentially coding region appeared to be interrupted by an intron and its sequence has allowed the construction of an evolutionary tree including Tuber magnatum and three different fungal species.

Keywords

Codon Agarose Polypeptide Assimilation dNTP 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Altschul, S. F., Gish, W., Miller, W. and Myers, E. W., 1990, Basic Local Alignment Tool, J. of Mol Biol 215: 403–410.Google Scholar
  2. Boguski, M. S., Lowe, T. M. J. and Tolstoshev, C. M., 1993, dbEST-database for “expressed sequence tags”, Nature Genetics 4: 332–333.PubMedCrossRefGoogle Scholar
  3. Bork, P., Ouzounis, C., Sander, C., Scharf, M., Schnaeider, R. and Sonnhamer, E., 1992, Comprehensive sequence analysis of the 182 predicted open reading frames of yeast chromosome III, Protein Sci. 1: 1677–1690.PubMedCrossRefGoogle Scholar
  4. Calabrò, V., Mason, P. J., Filosa, S., Civitelli, D., Cittadella, R., Tagarelli, A., Martini, G., Brancati, C. and Luzzatto, L., 1993, Genetic heterogeneity of glucose 6-phosphate dehydrogenase deficiency revealed by single-strand conformation and sequence analysis, Am. J. Hum. Genet. 52: 527–536.PubMedGoogle Scholar
  5. Devereux, J., Haeberli, P. and Smithies, O., 1984, A comprehensive set of sequence analysis program for the VAX, Nucleic Acids Research 12(1): 387–395.PubMedCrossRefGoogle Scholar
  6. Dujon, B. and the European Consortium for the Yeast Genome, 1994, Complete DNA sequence of Yeast chromosome XI, Nature 369: 371–378.PubMedCrossRefGoogle Scholar
  7. Filosa, S., Calabrò, V., Lania, G., Vulliamy, T. J., Brancati, C., Tagarelli, A., Luzzato, L. and Martini, G., 1993, G6PD haplotypes spanning Xq28 fromF8C to Red/Green color vision, Genomics 17:6–14.PubMedCrossRefGoogle Scholar
  8. Green, P., Lipman, D., Hillier, L., Waterston, R., States, D. and Claverie, J.-M., 1993, Ancient Conserved Regions in New Gene Sequences and the Protein Databases, Science 259: 1711–1716.PubMedCrossRefGoogle Scholar
  9. Green, E. D. and Olson, M. V., 1990, Systematic screening of yeast artificial-chromosome libraries by use of the polymerase chain reaction, Proc. Natl. Acad. Sci. USA 87: 1213–1217.PubMedCrossRefGoogle Scholar
  10. Habib Mohamed, A., Chirala, S., Mody, N. H., Hang, W.-Y and Wakil, S. J., 1988, Primary Structure of the Multifunctional alpha Subunit Protein of Yeast Fatty Acid Synthase Derived from FAS 2 Gene Sequence, J. Biol. Chem. 263(25): 12315–12325.Google Scholar
  11. Jeffery, J., Persson, B., Wood, I., Bergman, T., Jeffery, R. and Jornvall, H., 1993, Glucose-6-phosphate dehydrogenase. Structure-function relationship and the Pichia jadinii enzyme structure, Eur. J. Biochem. 212:41–49.PubMedCrossRefGoogle Scholar
  12. Lanfranco, L., Wyss, P., Marzachì, L. and Bonfante, P., 1994, DNA probes for the identification of the ectomycorrhizal fungus Tuber magnatum Pico, FEMS Microbiol. Lett. 114: 245–252.CrossRefGoogle Scholar
  13. Martin, F. M. and Hilbert, J. L., 1991, Morphological, biochemical and molecular changes during ectomycor- rhiza development, Experientia 47: 321–331.CrossRefGoogle Scholar
  14. McCombie, W. R., Adams, M. D., Kelley, J. M., FitzGerald, M. G., Utterback, T. R., Khan, M., Dubnick, M., Kerlavage, A. R., Venter, J. C. and Fields, C., 1992, Caenorhabditis elegans expressed sequence tags identify gene families and potential disease gene homologues., Nat. Genet 1: 124–31.PubMedCrossRefGoogle Scholar
  15. Mischiati, P., and Fontana, A. 1993, In vitro culture of Tuber magnatum mycelium isolated from mycorrhizas, Mycol Res. 97(1): 40–44.CrossRefGoogle Scholar
  16. O’Brien, E., Kurdi-Haidar, B., Wanachiwanawin, W., Carvajal, J. L., Vulliamy, T., Cappadoro, M., Mason, P. J. and Luzzatto, L., 1994, Cloning of the glucose 6-phosphate dehydrogenase gene from Plasmodium falciparum, Mol. Biochem. Parasitol. 64: 313–326.PubMedCrossRefGoogle Scholar
  17. Oliver, S. G. et al., 1992, The complete DNA sequence of yeast chromosome III, Nature 357: 38–46.PubMedCrossRefGoogle Scholar
  18. Rice, C. M., Fuchs, R., Higgins, D., Stoehr, P. J., and Cameron, G. N. 1993, The EMBL data library, Nucleic Acids Research 21: 2967–2971.PubMedCrossRefGoogle Scholar
  19. Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989). Molecular Cloning, a laboratory manual. Cold Spring Harbor, Cold Spring Harbor Laboratory Press.Google Scholar
  20. Schuller, H. J., Hahn, A., Troester, F., Schutz, A., Schweizer, M. and Schweizer, E., 1992, Coordinate genetic control of yeast fatty acid synthase genes FAS 1 and FAS 2 by an upstream activation site common to genes involved in membrane lipid byosynthesis, The EMBO Journal 11: 107–114.PubMedGoogle Scholar
  21. Schuller, H. J., Schutz, A., Knab, S., Hoffmann, S. and Schweizer, E., 1994, Importance of general regulatory factors Raplp, Abflp, and Reblp for the activation of yeast fatthy acid synthase genes FAS 1 and FAS 2, Eur. J. Biochem. 225: 213–222.Google Scholar
  22. Vulliamy, T., Mason, P. and Luzzatto, L., 1992, The molecular basis of glucose-6-phosphate dehydrogenase deficiency, Trends in Genetics 8(4): 138–143.PubMedCrossRefGoogle Scholar
  23. Weber, J. L., 1990, Informativeness of human (dC-dA)n-(dG-dT)n polymorphisms, Genomics 7: 524–530.PubMedCrossRefGoogle Scholar
  24. Wiesner, P., Beck, J., Beck, K.-F., Ripka, S., Muller, G., Lucke, S. and Schweizer, E., 1988, Isolation and sequence analysis of the fatty acid synthetase FAS2 gene from Penicillium patulum., Eur. J. Biochem. 177: 69–79.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • M. G. De Santo
    • 1
  • S. Filosa
    • 1
  • A. Franzè
    • 1
  • G. Martini
    • 1
  1. 1.Istituto Internazionale di Genetica e BiofisicaCNRNapoliItaly

Personalised recommendations