Suppression of tandem-multimer formation during genetic transformation of the mycotoxin-producing fungus Penicillium paxilli by disrupting an orthologue of Aspergillus nidulans uvsC

Abstract

An orthologue of Aspergillus nidulans uvsC and Saccharomyces cerevisiae RAD51 was cloned from the filamentous fungus, Penicillium paxilli. A mutation in uvsC causes UV sensitivity during germination. The product of RAD51 is involved in meiotic recombination and DNA damage repair. The deduced amino acid sequence of the product of this gene (Pprad51) shared 92% identity with UVSC. Site-specific disruption of pprad51 showed a significant effect for extra-cellular DNA integration. Transformation of the null mutant with pII99, which confers geneticin resistance, resulted in a shift from a predominance of direct repeats at a single site to single copies when compared with a control strain. A copy-number effect of integrated pII99 for geneticin selection was suggested as the frequency of direct repeat formation was less when selected at a lower concentration in the control strain. However, such an effect was not observed in the null mutant, further supporting an involvement of Pprad51 in direct repeat formation.