Current Genetics

, Volume 14, Issue 5, pp 451–456 | Cite as

Transformation as a method of increasing gene copy number and gene expression in the basidiomycete fungus Coprinus cinereuss

  • Fiona M. Mellon
  • Lorna A. Casselton
Original Articles
Received:

Summary

Seven transformants having varying numbers of non-homologously integrated copies of the isocitrate lyase gene, acu-7, were analysed for enzyme activity. Maximum levels of activity, 3.8 times that of the wild type, were observed in a transformant with only two gene copies whereas eight gene copies in another transformant led to only 25% wild type activity. Acu+ transformants were not selected directly for expression of acu-7 but as cotransformants. Analysis of 14 transformants not expressing acu-7 showed that four contained transforming DNA sequences and significantly, two had evidence of non-homologously integrated tandem duplications of the entire acu-7 plasmid DNA. The site of integration of the gene was thus important in determining whether or not it was. expressed and to what level it was expressed. A comparison of induced and uninduced levels of enzyme activity confirmed that the enzyme was still tightly regulated.

Key words

Isocitrate lyase Coprinus Transformation Gene expression 
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Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Fiona M. Mellon
    • 1
  • Lorna A. Casselton
    • 1
  1. 1.School of Biological SciencesQueen Mary CollegeLondonUK

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