Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 73, Issue 6, Pages 1348-1354Publisher
SPRINGER
DOI: 10.1007/s00253-006-0603-3
Keywords
Agrobacterium-mediated transformation; Trichoderma reesei; T-DNA; insertional mutagenesis; hygromycin B resistance; TAIL-PCR
Categories
Ask authors/readers for more resources
Filamentous fungus Trichoderma reesei QM9414 was successfully transformed with Agrobacterium tumefaciens AGL-1 for random integration of transforming DNA (T-DNA). Co-cultivation of T. reesei conidia or protoplasts with A. tumefaciens in the presence of acetosyringone resulted in the formation of hygromycin B-resistant fungal colonies with high transformation frequency. Nine randomly selected resistant clones were proved to be stable through mitotic cell division. The integration of the hph gene into T. reesei genome was determined by PCR and dot blot analysis. Transgenic T. reesei strains were analyzed using TAIL-PCR for their T-DNA contents. The results showed that T-DNA inserts occurred evidently by fusing DNA at T-DNA borders via random recombination, which suggests that Agrobacterium-mediated transformation is a potentially powerful tool towards tagged mutagenesis and gene transfer technology for T. reesei.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available