An efficient tool for random insertional mutagenesis: Agrobacterium tumefaciens-mediated transformation of the filamentous fungus Aspergillus terreus

Abstract

Agrobacterium tumefaciens-mediated transformation (ATMT) has been widely used in various organisms. In the current study, we developed a simple and efficient system for genetic transformation of the filamentous fungus Aspergillus terreus using ATMT. The transformation protocol was optimized for certain parameters to rapidly generate a library of Transferred DNA (T-DNA) insertion mutants of A. terreus. The presence of mitotically stable hygromycin resistance gene (hph) integration in the genome was confirmed by PCR, and T-DNA flanking sequences were cloned by thermal asymmetric interlaced PCR. The successful construction of the mutant library demonstrated the utility of the ATMT approach for future forward and reverse genetic studies in this important fungus.

Keywords: Agrobacterium tumefaciens; Aspergillus terreus; Filamentous fungus; T-DNA flanking sequences; Transformation.