[Transformation system of Aspergillus japonicus mediated by Agrobacterium tumefaciens]

In to order to select Aspergillus japonicus mutant strains that express high-yield of glycerol oxidase, we constructed a mutant library of Aspergillus japonicus that mediated by Agrobacterium tumefaciens.

Tri-relative hybridization experiments were performed to transfer binary vector pB I -hph II into Agrobacterium tumefaciens EHA105 that was used as the infective strain and Aspergillus japonicus was used as the host strain. We established the transformation system of Aspergillus japonicus mediated by Agrobacterium tumefaciens and constructed a mutant library. At the same time, we analyzed the factors affecting the transformation efficiency such as the concentration of Agrobacterium tumefaciens, adding and not adding Acetosyringone, co-culture time, and co-culture temperature.

PCR detection and Southern blot analysis showed that the T-DNA was integrated into genme of the Aspergillus japonicus. All 9 randomly selected transformants were stable after 10 rounds of successive cultures.

The transformation system is a good basis for selection of Aspergillus japonicus mutant strains that express high-yield of glycerol oxidase.