Transformation of a nitrate reductase deficient mutant of Penicillium chrysogenum with the corresponding Aspergillus niger and A. nidulans niaD genes |
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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 |
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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 |
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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. |
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Keywords: | Penicillium chrysogenum Nitrate reductase niaD mutants Nitrate assimilation Fungal transformation |
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