Skip to main content
Log in

Functional elements of the promoter region of the Aspergillus oryzae glaA gene encoding glucoamylase

  • Original Articles
  • Published:
Current Genetics Aims and scope Submit manuscript

Summary

Analysis was made of the promoter region of the Aspergillus oryzae glaA gene encoding glucoamylase. Northern blots using a glucoamylase cDNA as a probe indicated that the amount of mRNA corresponding to the glaA gene increased when expression was induced by starch or maltose. The promoter region of the glaA gene was fused to the Escherichia coli uidA gene, encoding β-glucuronidase (GUS), and the resultant plasmid was introduced into A. oryzae. Expression of GUS protein in the A. oryzae transformants was induced by maltose, indicating that the glaA-GUS gene was regulated at the level of transcription in the presence of maltose. The nucleotide sequence 1.1 kb upstream of the glaA coding region was determined. A comparison of the nucleotide sequence of the A. oryzae glaA promoter with those of A. oryzae amyB, encoding α-amylase, and A. niger glaA showed two regions with similar sequences. Deletion and site-specific mutation analysis of these homologous regions indicated that both are essential for direct high-level expression when grown on maltose.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Caruthers MH (1982) Chemical synthesis of oligonucleotides using the phosphite triester intermediates. In: Gassen HG, Lang A (eds) Chemical and enzymatic synthesis of gene fragments. A laboratory manual. Verg. Chemie, Weinheim, pp 71–79

    Google Scholar 

  • Cathala G, Savouret JF, Mandez B, West BL, Karin M, Martial JA, Baxter JD (1983) DNA 2:329–335.

    Google Scholar 

  • Christensen T, Woeldike H, Boel E, Mortensen SD, Hjortshoej K, Thim L, Hansen MT (1988) Bio/Technology 6:1419–1422

    Google Scholar 

  • El-Goagary S, Leite A, Crivellaro O, Eveleigh DE, El-Dorry H (1989) Proc Natl Acad Sci USA 86:6138–6141.

    Google Scholar 

  • Fowler T, Berka M, Ward M (1990) Curr Genet 18:537–545

    Google Scholar 

  • Gomi K, Iimura Y, Hara S (1987) Agric Biol Chem 51:2549–2555

    Google Scholar 

  • Hamer JE, Timberlake WE (1987) Mol Cell Biol 7:2352–2359

    Google Scholar 

  • Hanahan D (1983) J Mol Biol 166:557–580

    Google Scholar 

  • Hata Y, Kitamoto K, Gomi K, Kumagai C, Tamura G, Hara S (1991a) Agric Biol Chem 55:941–949

    Google Scholar 

  • Hata Y, Tsuchiya K, Kitamoto K, Gomi K, Kumagai C, Tamura G, Hara S (1991b) Gene 108:145–150

    Google Scholar 

  • Higuchi R (1989) Using PCR to engineer DNA. In: Erlich HA (ed) PCR Technology. Stockton Press, New York, pp 61–70

    Google Scholar 

  • Iimura Y, Gomi K, Uzu H, Hara S (1987) Agric Biol Chem 51:323–328

    Google Scholar 

  • Jefferson RA, Burgess SM, Hirsh D (1986) Proc Natl Acad Sci USA 83:8447–8451

    Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) EMBO J 6:3901–3907

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Fan AL, Randall RJ (1951) J Biol Chem 193:268–275

    Google Scholar 

  • Punt PJ, Dingemanse MA, Kuyvenhoven A, Soede RDM, Pouwels PH, Van den Hondel CAMJJ (1990) Gene 93:101–109

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. 1989: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) Proc Acad Natl Sci USA 74:5463–5467

    Google Scholar 

  • Struhl K (1987) Cell 49:295–297

    Google Scholar 

  • Tada S, Iimura Y, Gomi K, takahashi K, Hara S, Yoshizawa K (1989) Agric Biol Chem 53:593–399

    Google Scholar 

  • Tada S, Gomi K, Kitamoto K, Kumagai C, Tamura G, Hara S (1991a) Agric Biol Chem 55:1939–1941

    Google Scholar 

  • Tada S, Gomi K, Kitamoto K, Takahashi K, Tamura G, Hara S (1991b) Mol Gen Genet 229:301–306

    Google Scholar 

  • Unkles SE, Campbell EI, de Ruiter-Jacobs YMJT, Broekhuijsen M, Marco JA, Carrez D, Contreras R, Van den Hondel CAMJJ, Kinghorn JR (1989) Mol Gen Genet 218:99–104

    Google Scholar 

  • Vieira and Messing (1987) Method Enzymol 153:3–11

    Google Scholar 

  • Ward M, Wilson LJ, Kodama KH, Rey MW, Berka RM (1990) Bio/Technology 8:435–438

    Google Scholar 

  • Wirsel S, Lachmund A, Wildhardt G, Ruttkowski E (1989) Mol Microbiol 3:3–14

    Google Scholar 

  • Yanish-Perron C, Vieira J, Messing J (1985) Gene 33:103–119

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by M. Yamamoto

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hata, Y., Kitamoto, K., Gomi, K. et al. Functional elements of the promoter region of the Aspergillus oryzae glaA gene encoding glucoamylase. Curr Genet 22, 85–91 (1992). https://doi.org/10.1007/BF00351466

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00351466

Key words

Navigation