Establishment of the transformation system of Aspergillus japonicus mediated by Agrobacterium tumefaciens

Abstract:

Abstract: [Objective]n 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. [Methods]Tri-relative hybridization experiments were performed to transfer binary vector pBI-hphII into Agrobacterium tumefaciens EHA105, which 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, co-culture temperature and so on. [Results]The results of PCR detection and Southern analysis showed that the T-DNA was integrated into genome of the Aspergillus japonicus. All of the nine randomly selected transformants were quite stable after ten rounds of successive cultures. [Conclusion]The transformation system is a good basis for selection of Aspergillus japonicus mutant strains which give a high-yield of glycerol oxidase.