Abstract
The brlA and abaA genes of Aspergillus nidulans regulate stages of conidiophore development and are themselves regulated during development. brlA − mutants produce conidiophore stalks devoid of vesicles, sterigmata, and spores. abaA − mutants produce most of the conidiophore structures but fail to form conidia. To assess the spatial expression of these two genes, we fused the 5′ flanking region of brlA or abaA to the Escherichia coli lacZ gene. A. nidulans transformants with a single copy of either fusion gene integrated at a defined heterologus locus ( argB) expressedβ-galactosidase during conidiophore development, paralleling brlA and abaA mRNA accumulation. Controls lacking the fusion genes produced little or noβ-galactosidase activity. A method for in situ detection ofβ-galactosidase was devised. Hyphae or conidiophores were permeabilized by treatment with chloroform vapors and stained with 5-bromo-4-chloroindolyl-β- d-galactoside.β-Galactosidase activity was detected in specific conidiophore cell types. brlA- and abaA-directedβ-galactosidase accumulated in vesicles, sterigmata, and immature conidia. This procedure should be applicable for determining cellular specificities of gene expression in fungi for which transformation systems exist.
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