Heterologous expression of the immunomodulatory protein gene from Ganoderma sinense in the basidiomycete Coprinopsis cinerea.
OBJECTIVE
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.


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 314mg kg(-1) fresh mycelia. The molecular weight of the FIP-gsi was 13kDa, 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.


CONCLUSIONS
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.
Han
F
F
Department of Bioengineering, College of Food Science, South China Agricultural University, Guangzhou, China.
Liu
Y
Y
Guo
L Q
LQ
Zeng
X L
XL
Liu
Z M
ZM
Lin
J F
JF
D000818:Animals; D001487:Basidiomycota / Q000235:genetics*; D001709:Biotechnology; D015152:Blotting, Northern; D015139:Blotting, Southern; D005656:Fungal Proteins / Q000235:genetics* / Q000302:isolation & purification; D032902:Ganoderma / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005822:Genetic Vectors / Q000235:genetics; D006386:Hemagglutination Tests; D006801:Humans; D007155:Immunologic Factors / Q000235:genetics; D051379:Mice; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014170:Transformation, Genetic
D005656:Fungal Proteins; D007155:Immunologic Factors