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Transformation of a nitrate reductase deficient mutant of Penicillium chrysogenum with the corresponding Aspergillus niger and A. nidulans niaD genes
Authors:Michael P. Whitehead   Shiela E. Unkles   Martin Ramsden   Edward I. Campbell   Sarah Jane Gurr   David Spence   Cees van den Hondel   Roland Contreras  James R. Kinghorn
Affiliation:(1) Plant Molecular Genetics Unit, Sir Harold Mitchell Building, Greenside Place, KY16 9TH St. Andrews, Fife, UK;(2) Genetic Laboratory, Glaxochem Ltd., LA12 9DR Ulverston, Cumbria, UK;(3) Medical Biological Laboratory TNO, P.O. Box 45, 2280 AA Rijswijk, The Netherlands;(4) Laboratorium voor Moleculaire Biologie, University of Gent, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium
Abstract:Summary A heterologous gene mediated transformation system based on niaD, the structural gene encoding nitrate reductase, has been developed for Penicillium chrysogenum. Transformation frequencies of up to 20 transformants per microgram DNA were obtained using the Aspergillus nidulans gene and 9 transformants per microgram using the A. niger gene. Vector constructs carrying the A. nidulans ans-1 sequence and the A. niger niaD gene did not show increased transformation frequencies. Southern blot hybridisation analysis demonstrated that vector sequences had integrated into the recipient genome. The control of heterologous niaD gene expression generally agreed with that found in the wild-type strain, that is, induction by nitrate and repression in the presence of ammonium.
Keywords:Penicillium chrysogenum  Nitrate reductase  niaD mutants  Nitrate assimilation  Fungal transformation
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