Improved transformation efficiency of Aspergillus niger using the homologous niaD gene for nitrate reductase |
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| Authors: | Edward I. Campbell Shiela E. Unkles Janet A. Macro Cees van den Hondel Roland Contreras James R. Kinghorn |
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| Affiliation: | (1) Plant Molecular Genetics Unit, Sir Harold Mitchell Building, University of St. Andrews, KY16 9TH St. Andrews, Fife, UK;(2) Medical Biological Laboratory TNO, P.O. Box 45, NL-2280 AA Rijswijk, The Netherlands;(3) Laboratorium voor Moleculaire Biologie, University of Gent, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium |
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| Abstract: | Summary Aspergillus niger transformation frequencies of up to 1,176 transformants per  g DNA were achieved using the plasmid vector pSTA10 containing the A. niger nitrate reductase structural gene. Analysis of genomic endonuclease cleaved DNA from nitrate utilising transformants by DNA hybridisation, showed that most integration events are as a result of homologous recombination. The niaD transformation system was used successfully for the introduction of the unselected Escherichia coli fusion genes lacZ, encoding  -galactosidase, and uidA, for -glucuronidase, as well as the Neurospora crassa tub-2 gene, for -tubulin. pSTA10 was also capable of transforming niaD mutants of other filamentous fungi such as A. nidulans, A. oryzae and Penicillium chrysogenum. |
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| Keywords: | Aspergillus niger Nitrate reductase Transformation of fungi Integration of plasmids Co-transformation frequencies |
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