Skip to main content
SpringerLink
Log in

cDNA cloning and expression of aTalaromyces emersonii amylase encoding genetic determinant inEscherichia coli

  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Summary

Intact mRNA has been isolated from the thermophilic fungusTalaromyces emersonii following growth on starch containing media. This has been used as template to synthesise cDNA. The cDNA was cloned into theEscherichia coli expression vector system, pUC18 and this was used to transformE. coli. Transformed colonies were screened for production of amylase activity and a number of positive recombinants were detected. One of those was found to contain a plasmid named pMH1, which harboured a 1.2 kb insert. Sub-cloning experiments verified that the amylase phenotype was encoded for by this fragment. The fragment was characterised using restriction enzyme cleavage site analysis. The origin of the insert was verified using both Northem and Southem blotting hybridisation analysis ofT. emersonii RNA and DNA respectively.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bethesda Research Labs. (1984) Focus, 6.

  • Bunni,L. & McHale,A.P. (1989) Biotechnol. Techniques Vol.3, 2, 107–112.

    Google Scholar 

  • Bunni,L., McHale,L. & McHale,A.P. (1989) Enz. Microb. Technol. Vol. 11, 370–375.

    Google Scholar 

  • Folan,M. & Coughlan,M.P. (1981) Int. J. Biochem. Vol. 13, 243–245.

    Google Scholar 

  • Hendy,L., Gallagher,J., Winters,A., Hackett,T.J., McHale,L. & McHale, A.P. (1990) Biotechnol. Letts. Vol. 12, 9, 673–678.

    Google Scholar 

  • Itkar,P. & Tsukagashi,N. (1990) Biochem. Biophys. Res. Comm. Vol. 166, 630–636.

    Google Scholar 

  • Maniatis,T., Fritsch,E. & Sambrook,J. (1982) Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory.

    Google Scholar 

  • McHale,A.P. & Coughlan,M.P. (1981) Biochim. Biophys. Acta., Vol.662 145–151.

    Google Scholar 

  • Nakakima,R., Imanaka,T. & Aiba,S. (1986) Appl. Microbiol. Biotechnol. Vol. 23, 255–360.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Dept. of Biological and Biomedical Sciences, University of Ulster, BT52 1SA, Coleraine, Northem Ireland

    L. McHale, T. J. Hackett & A. P. McHale

  2. Dept. of Dental Health, Trinity College Dublin, Ireland

    D. C. Coleman

  3. A.Anderson Consulting, London, UK

    L. Bunni

Authors
  1. L. Bunni
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. C. Coleman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L. McHale
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T. J. Hackett
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. P. McHale
    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

Bunni, L., Coleman, D.C., McHale, L. et al. cDNA cloning and expression of aTalaromyces emersonii amylase encoding genetic determinant inEscherichia coli . Biotechnol Lett 14, 1109–1114 (1992). https://doi.org/10.1007/BF01027011

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01027011

Keywords

Search

Navigation