Journal of Applied Microbiology, Vol.109, No.5, 1838-1844, 2010
Heterologous expression of the immunomodulatory protein gene from Ganoderma sinense in the basidiomycete Coprinopsis cinerea
Aims: FIP-gsi, a fungal immunomodulatory protein found in Ganoderma sinense, has antitumour, anti-allergy and immunomodulatory activities and is regulated by the fip-gsi gene. In this study, we aimed to express the fip-gsi gene from G. sinense in Coprinopsis cinerea to increase yield of FIPs-gsi. Methods and Results: A fungal expression vector pBfip-gsi containing the gpd promoter from Agaricus bisporus and the fip-gsi gene from the G. sinense was constructed and transformed into C. cinerea. PCR and Southern blotting analysis verified the successful integration of the exogenous gene fip-gsi into the genome of C. cinerea. RT-PCR and Northern blotting analysis confirmed that the fip-gsi gene was transcribed in C. cinerea. The yield of the FIP-gsi protein reached 314 mg kg-1 fresh mycelia. The molecular weight of the FIP-gsi was 13 kDa, and the FIP-gsi was capable of hemagglutinating mouse red blood cells, but no such activity was observed towards human red blood cells in vitro. Conclusions: The fip-gsi from G. sinense has been successfully translated in C. cinerea, and the yield of bioactive FIP-gsi protein was high. Significance and Impact of the Study: This is the first report using the C. cinerea for the heterologous expression of FIP-gsi protein and it might supply a basis for large-scale production of the protein.
Keywords:Coprinopsis cinerea;fip-gsi gene;fungal immunomodulatory proteins;Ganoderma sinense;heterologous expression