Development of a homologous transformation system for Aspergillus niger based on the pyrG gene
| Autor: | W. van Hartingsveldt, P.H. Pouwels, C.A.M.J.J. van den Hondel, I. E. Mattern, C. M. J. Van Zeijl |
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| Rok vydání: | 1987 |
| Předmět: |
Genetics
biology Carboxy-Lyases fungi Mutant Aspergillus niger Genes Fungal Orotidine-5'-Phosphate Decarboxylase biology.organism_classification Neurospora crassa Transformation (genetics) Plasmid Transformation Genetic Cotransformation Genes Aspergillus nidulans Mutation Molecular Biology Orotidine 5'-phosphate decarboxylase Plasmids |
| Zdroj: | Moleculargeneral genetics : MGG. 206(1) |
| ISSN: | 0026-8925 |
| Popis: | The development of a homologous transformation system for Aspergillus niger is described. The system is based on the use of an orotidine-5'-phosphate decarboxylase deficient mutant (pyrG) and a vector, pAB4-1, which contains the functional A. niger pyrG gene as a selection marker. Transformation of the A. niger pyrG mutant with pAB4-1 resulted in the appearance of stable Pyr+ transformants at a frequency of 40 transformants per microgram of DNA. In 90% of these transformants integration had occurred at the resident pyrG locus, resulting either in replacement of the mutant allele by the wild-type allele (60%) or in insertion of one or two copies of the vector (40%). The A. niger pyrG mutant could also be transformed with the vector pDJB2 containing the pyr4 gene of Neurospora crassa, at a frequency of 2 transformants per microgram of DNA. Integration at the resident pyrG locus was not found with this vector. The vector pAB4-1 is also capable of transforming an Aspergillus nidulans pyrG mutant to Pyr+. The pyrG transformation system was used for the introduction of a non-selectable gene into A. niger. |
| Databáze: | OpenAIRE |
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