Development of host strains and vector system for an efficient genetic transformation of filamentous fungi

Publication date: Available online 19 November 2018Source: PlasmidAuthor(s): Larissa A. Balabanova, Yuri N. Shkryl, Lubov V. Slepchenko, Yulia A. Yugay, Tatiana Y. Gorpenchenko, Natalia N. Kirichuk, Yulia V. Khudyakova, Irina Yu. Bakunina, Anna B. Podvolotskaya, Victor P. Bulgakov, Aleksandra V. Seitkalieva, Oksana M. Son, Liudmila A. TekutyevaAbstractAn ability to synthesize extracellular enzymes degrading a wide spectrum of plant and algae polymeric substrates makes many fungi relevant for biotechnology. The terrestrial thermophilic and marine fungal isolates capable of plant and algae degradation have been tested for antibiotic resistance for their possible use in a new genetic transformation system. Plasmids encoding the hygromycin B phosphotransferase (hph) under the control of the cauliflower mosaic virus 35S promoter, the trpC gene promoter of Aspergillus nidulans, and the Aureobasidium pullulans TEF gene promoter were delivered into the fungal cells by electroporation. The effectiveness of different promoters was compared by transformation and growth of Thermothelomyces thermophila (formerly Myceliophthora thermophila) on the selective medium and by real-time PCR analysis. A highly efficient transformation was observed at an electric-pulse of 8.5 kV/cm by using 10 μg of DNA per 1 × 105 conidia. Although all promoters were capable of hph expression in the Th. thermophila cells, the trpC promoter provided the highest level of hygromycin resistance. We further ...
Source: Plasmid - Category: Biotechnology Source Type: research