NDLI: Increased transformation frequency and tagging of developmental genes in Aspergillus nidulans by restriction enzyme-mediated integration (REMI)

Content Provider	Springer Nature Link
Author	Sánchez, O. Navarro, R. E. Aguirre, J.
Copyright Year	1998
Abstract	We have used a plasmid containing the argB gene to transform an Aspergillus nidulansargB-deleted strain in the presence of restriction enzymes and show a 20- to 60-fold increase in transformation frequency via restriction enzyme-mediated integration (REMI). This procedure was used to try to tag new genes involved in the asexual development of this fungus. More than 2000 transformants isolated following electroporation of conidia and ∼3700 transformants recovered following protoplast fusion were screened for sporulation defects. Unexpectedly, developmental mutants were obtained only when the protoplast fusion approach was used. Southern blot analysis of these mutants, and of randomly selected transformants obtained by electroporation, was consistent with the occurrence of single plasmid integration events in 33 and 65% of the cases, respectively. The argB marker was shown to be tightly linked to the mutant phenotype in only 62% of the mutants analyzed by sexual crosses. Partial DNA sequencing of a tagged gene, whose mutation delays asexual sporulation and results in a fluffy phenotype, showed no homology to previously reported sequences. Our results indicate that REMI can be used in A. nidulans to increase the transformation frequency and illustrate the advantages and potential problems when using REMI to tag genes of interest in this and other fungi.
Starting Page	89
Ending Page	94
Page Count	6
File Format	PDF
ISSN	00268925
Journal	Molecular and General Genetics MGG
Volume Number	258
Issue Number	1
e-ISSN	14321874
Language	English
Publisher	Springer-Verlag
Publisher Date	1998-04-29
Publisher Place	Berlin/Heidelberg
Access Restriction	Subscribed
Content Type	Text
Resource Type	Article
Subject	Genetics

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