Isolation and sequence analysis of a beta-tubulin gene from Aspergillus flavus and its use as a selectable marker.

摘要: An altered beta-tubulin gene that confers resistance to benomyl [whose active ingredient is 2-(methoxycarbonylamino)benzimidazole (MBC)] was isolated from a DNA library of Aspergillus flavus and used as selectable marker for transformation. The cloned into plasmid vector containing the pyr-4 Neurospora crassa, transformants were selected either uracil prototrophy or MBC resistance. Transformants three phenotypic classes: sensitive, intermediate, resistant MBC. Transforming appeared integrate at several sites in genome, with more phenotypes having copies than sensitive intermediate phenotypes. also on medium average frequency transformation (1 3 per micrograms transforming DNA) lower obtained by selection prototrophy, presumably because failure select contained few gene. sequence determined compared published benA A. nidulans; found be highly conserved between two species. Notable differences lacks intron 6 present has an additional leucine position 148. This first reported aflatoxin-producing fungus adds growing body knowledge genes their use markers filamentous fungi.