Integrative and replicative transformation of Penicillium canescens with a heterologous nitrate-reductase gene

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 AMA 1 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.

Development of Transformation System of Verticillium lecanii (Lecanicillium spp.) (Deuteromycotina: Hyphomycetes) Based on Nitrate Reductase Gene of Aspergillus nidulans

“…Since the isolated mutants were spontaneous and because mutation is a random event, the number of isolated niaD -mutants for each species will predictably not follow a pattern, as reported for Paecilomyces griseoroseum, where 8.33% of the chlorate resistants mutants were niaD -mutants, while in P. canescens the isolated niaD -mutants made up 49.0% [20].…”

Using an Inducible Promoter of a Gene Encoding Penicillium verruculosum Glucoamylase for Production of Enzyme Preparations with Enhanced Cellulase Performance

“…verruculosum B1-537 strain jointly with a transforming plasmid pSTA10 (10:1, μg), using the modified method described by Aleksenko et al [26]. The pSTA10 plasmid contains a nitrate reductase gene providing complementation of a defective niaD gene in the host strain.…”

Glucoamylases from Penicillium verruculosum and Myceliophthora thermophila: Analysis of differences in activity against polymeric substrates based on 3D model structures of the intact enzymes

“…This allowed selection of the transformants on minimal media with NaNO 3 . Transformation efficiencies typically reach hundreds of transformants per mg of transforming DNA, with co-transformation frequencies of 80% and higher [13].…”

“…The pPrXylA-GA plasmid construct was directed into protoplasts of the recipient RN3-11-7 (niaD-) strain together with a plasmid pSTA10 (10:1, mg), using the modified method described by Aleksenko et al [13]. The pSTA10 plasmid, used for transformation, carried a nitrate reductase gene providing complementation of a defective niaD gene in the recipient strain.…”