Interaction between Fus3 and Ste12 in Aspergillus oryzae analysed by Bimolecular fluorescence complementation

Abstract:

[Background] Bimolecular fluorescence complementation (BiFC) is applied to explore the protein-protein interactions in some organisms such as Fusarium fujikuroi, but not yet applied in Aspergillus oryzae. [Objective] We used BiFC system to detect the interactions between proteins Fus3 and Ste12 in Aspergillus oryzae during growth and development. We used this system to specifically visualize the location of the target proteins in living cells. [Methods] Complex vectors were constructer by using the Multisite Gateway cloning system. Proteins Fus3 and Ste12 were fused to C- and N-terminal green fluorescence protein fragments, respectively, and then the hyphae of the obtained transformants were observed under a fluorescence microscope. [Results] The green fluorescence protein was observed in successfully transformed cells, indicating that the interaction occurred between Fus3 and Ste12 happened in A. oryzae. [Conclusion] The method to detect protein-protein interaction in A. oryzae proves its broader application possibility.