Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

Characterization of Two Cel (Cellulose Degradation) Genes of Clostridium Thermocellum Coding for Endoglucanases

Bio/Technology volume 1, pages 589–594 (1983)Cite this article

Abstract

Two structural genes, celA and celB, coding respectively for the endoglucanases A and B of Clostridium thermocellum, were subcloned from cosmids of a gene bank into plasmid pBR322. Both genes are carried in a DNA sequence of about 2.7 kb. They share no homology and are not contiguous on the C. thermocellum chromosome. They are expressed in E. coli independently of their orientation in the cloning vector.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Ng, T.K., Weimer, P.J. and Zeikus, J.G. 1977. Cellulolytic and physiological properties of Cloitridium thermocellum. Arch. Microbiol. 114: 1–7.

    Article  CAS  PubMed  Google Scholar 

  2. Zeikus, J.G. 1980. Chemical and fuel production by anaerobic bacteria. Annu. Rev. Microbiol. 34: 423–464.

    Article  CAS  PubMed  Google Scholar 

  3. Aït, N., Creuzet, N. and Forget, P. 1979. Partial purification of cellulase from Clostridium thermocellum. J. Gen. Microbiol. 113: 399–402.

    Article  Google Scholar 

  4. Shinmyo, A., Garcia-Mardnez, D.V. and Demain, A.L. 1979. Studies on the extracellular cellulolytic enzyme complex produced by Clostridium thermocellum. 1979. J. Appl. Biochem. 1: 202–209.

    CAS  Google Scholar 

  5. Garcia-Martinez, D.V., Shinmyo, A., Madia, A. and Demain, A.L. 1980. Studies on the cellulase produced by Clostridium thermocellum. European J. Appl. Microbiol. Biotechnol. 9: 189–197.

    Article  CAS  Google Scholar 

  6. Petre, J., Longin, R. and Millet, J. 1981. Purification and properties of an endo-β-1,4-glucanase from Clostridium thermocellum. Biochimie 63: 629–639.

    Article  CAS  PubMed  Google Scholar 

  7. Ng, T.K. and Zeikus, J.G. 1981. Purification and characterization of an endoglucanase (1,4-β-D-glucan glucanohydrolase) from Clostridium thermocellum. Biochem. J. 199: 341–350.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Creuzet, N. and Frixon, C. 1983. Purification and characterization of an endoglucanase from a newly isolated thermophilic anaerobic bacterium. Biochimie 65: 149–156.

    Article  CAS  PubMed  Google Scholar 

  9. Aït, N., Creuzet, N. and Cattaneo, J. 1982. Properties of β-glucosidase purified from Clostridium thermocellum. J. Gen. Microbiol. 128: 569–577.

    Google Scholar 

  10. Cornet, P., Tronik, D., Millet, J. and Aubert, J.-P. 1983. Cloning and expression in Escherichia coli of Clostridium thermocellum genes coding for amino acid synthesis and cellulose hydrolysis. FEMS Microbiol. Letters 16: 137–141.

    Article  CAS  Google Scholar 

  11. Teather, R.M. and Wood, P.J. 1982. Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Appl. Environ. Microbiol. 43: 777–780.

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Uhlin, B.E. and Nordstrom, K. 1977. R plasmid gene dosage effects in Escherichia coli K-12: copy mutants of the R plasmid RIdrd-19. Plasmid 1: 1–7.

    Article  CAS  PubMed  Google Scholar 

  13. Whittle, D.J., Kilburn, D.G., Warren, R.A.J. and Miller, R.C. Jr., 1982. Molecular cloning of a Cellulomonas fimi cellulase gene in Escherichia coli. Gene 17: 139–145.

    Article  CAS  PubMed  Google Scholar 

  14. Collmer, A. and Wilson, D.B. 1983. Cloning and Expression of a Thermonumospora YX Endocellulase Gene in E. coli. BIO/TECHNOLOGY 1: 594–601.

    CAS  Google Scholar 

  15. Boyer, H.W. and Roulland-Dussoix, D. 1969. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J. Mol. Biol. 41: 459–472.

    Article  CAS  PubMed  Google Scholar 

  16. Reeve, J.N. 1977. Bacteriophage infection of minicells. A general method for identification of in vivo bacteriophage directed polypeptide biosynthesis. Mol. Gen. Genet. 158: 73–79.

    Article  CAS  PubMed  Google Scholar 

  17. Bolivar, F., Rodriguez, R.L., Greene, J.P., Betlach, M.C., Heyneker, H.L. and Boyer, H.W. 1977. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2: 95–113.

    Article  CAS  PubMed  Google Scholar 

  18. Miller, J.H. 1972. In Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

    Google Scholar 

  19. Humphreys, G.O., Willshaw, G.A. and Anderson, E.S. 1975. A simple method for the preparation of large quantities of pure plasmid DNA. Biochim. Biophys. Acta 382: 457–463.

    Article  Google Scholar 

  20. Birnboim, H.C. and Doly, J. 1979. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 7: 1513–1523.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning, a Laboratory Manual. Cold Spring Harbor Laboratory, New York.

  22. Cohen, S.N., Chang, A.C.Y. and Hsu, L. 1972. Non chromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc. Natl. Acad. Sci. (USA) 69: 2110–2114.

    Article  CAS  Google Scholar 

  23. Rigby, P.W.J., Dieckmann, M., Rhodes, C. and Berg, P. 1977. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J. Mol. Biol. 113: 237–251.

    Article  CAS  PubMed  Google Scholar 

  24. Southern, E. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98: 503–517.

    Article  CAS  PubMed  Google Scholar 

  25. Quiviger, B., Franche, C., Lutfalla, G., Rice, D., Haselkorn, R. and Elmerich, C. 1982. Cloning of a nitrogen-fixation (nif) gene cluster of Azaspirillum brasilense. Biochimie 64: 495–502.

    Article  CAS  PubMed  Google Scholar 

  26. Kessler, S.W. 1976. Cell membrane antigen isolation with the staphylococcal protein A-antibody adsorbent. J. Immunol. 117: 1482–1496.

    CAS  PubMed  Google Scholar 

  27. Béguin, P., Nicolas, P., Boussetta, H., Fahy, C. and Cohen, P. 1981. Characterization of the 80,000 molecular weight form of neurophysin isolated from bovine neurohypophysis. J. Biol. Chem. 256: 9289–9294.

    PubMed  Google Scholar 

  28. Pardee, A.B., Jacob, F. and Monod, J. 1959. The genetic control and cytoplasmic expression of “inducibility” in the synthesis of β-galactosidase by Escherichia coli. J. Mol. Biol. 1: 165–178.

    Article  CAS  Google Scholar 

  29. Ross, G.W. and O'Callaghan, C.H. 1975. β-lactamase assays. In Methods in Enzymology, J. H. Hash (ed.), Academic Press, New York 43: 69–85.

    Google Scholar 

  30. Heppel, L.A. 1967. Selective release of enzymes from bacteria. Science 156: 1451–1455.

    Article  CAS  PubMed  Google Scholar 

  31. Sedmak, J.J. and Grossberg, S.E. 1977. A rapid, sensitive and versatile assay for protein using Coomassie brilliant blue G250. Anal. Biochem. 79: 544–552.

    Article  CAS  PubMed  Google Scholar 

  32. Lammli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Article  Google Scholar 

Download references

Author information

Author notes

  1. Philippe Cornet: On leave from Solvay et Cie, Brussels, Belgium.

Authors and Affiliations

  1. Unité de Physiologie Cellulaire, Département de Biochimie et Génétique Moléculaire, Institut Pasteur, 75724, Paris, Cedex 15, France

    Philippe Cornet, Jacqueline Millet, Pierre Béguin & Jean-Paul Aubert

Authors
  1. Philippe Cornet
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jacqueline Millet
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pierre Béguin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jean-Paul Aubert
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cornet, P., Millet, J., Béguin, P. et al. Characterization of Two Cel (Cellulose Degradation) Genes of Clostridium Thermocellum Coding for Endoglucanases. Nat Biotechnol 1, 589–594 (1983). https://doi.org/10.1038/nbt0983-589

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0983-589

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing