Integrative and replicative transformation of Penicillium canescens with a heterologous nitrate-reductase gene |
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| Authors: | A. Y. Aleksenko N. A. Makarova I. V. Nikolaev A. J. Clutterbuck |
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| Affiliation: | (1) Genetics Laboratories, Institute of Biological and Life Sciences, University of Glasgow, G11 5JS Glasgow, Scotland, UK;(2) Institute of Genetics and Selection of Industrial Microorganisms, 1st Dorozhny 1, 113545 Moscow, Russia |
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| Abstract: | A wild isolate of Penicillium canescens was subjected to mutagenesis, and 150 chlorate-resistant mutants were isolated and classified in respect of their ability to utilize various nitrogen sources. Strains supposedly deficient in nitrate reductase have been transformed with the nitrate-reductase gene from Aspergillus niger. Transformation probably occurred by non-homologous integration of the transforming vector into the chromosome. Co-transformation with the AMA1 replicating element from A. nidulans enhanced transformation frequency up to 2000-fold, and was shown to result in autonomous maintenance of replicating concatenates, one of which was re-isolated by transformation of E. coli. |
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| Keywords: | AMA1 Chlorate-resistance Co-transformation niaD gene |
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