Tagging of genes that confer multidrug resistance in Aspergillus nidulans by restriction enzyme-mediated integration (REMI).

Gustavo H. Goldman, Cristiane C. de Souza, and Maria Helena S. Goldman.

Abstract: 

As a preliminary step to characterizing genes that confer pleiotropic drug resistance in Aspergillus nidulans, we isolate transformants by REMI (Restriction Enzyme-Mediated Integration) that show pleiotropic drug sensitivity. We have used a plasmid containing the pyr4 gene to transform an A. nidulans pyre mutant in the presence of BamHI. One thousand two-hundred sixty-seven transformants were isolated using the plasmid pRG3 digested with BamHI. Southern analysis of these mutants, and of randomly selected transformants was consistent with the occurrence of single plasmid integration events in about 70 % of the cases. Approximately 900 of these transformants were examined for sensitivity to fourty-seven drugs or stress agents with different and/or the same mechanism of action. Thirty-five transformants displayed sensitivity to a single drug (either itraconazole, miconazole, hygromycin or cycloheximide) while six of them displayed multidrug-sensivity. The pyr4 marker was shown to be tightly linked to the mutant phenotype in only one from these multidrug-sensitivity transformants. Financial support: FAPESP and CNPq, Brazil, and ICGEB-UNIDO
1999

abstract No: 

Fungal Genet. Newsl. 46 (Supl):

Full conference title: 

Fungal Genetics Conference 20th
    • Fungal Genetics Conference 20th (1999)