Diabetes is a serious, chronic disease that occurs either when the human body does not produce enough insulin, or when the body cannot effectively use the insulin it produces. The stages of protein metabolism in mushrooms are very similar to those in animal cells and plants. As a result, mushrooms have high potential to be employed in production of complex proteins such as secretion antibodies and oral vaccines. The importance of edible mushrooms is not only limited to their nutritional properties as delicious foodstuff, but also they play roles as food supplements, pharmaceuticals and bioremediation properties and their ability for production of recombinant proteins. Among of all known cultivated mushrooms, the white button mushroom, Agaricus bisporus, hasthe ability to make disulfide bonds and the post-translational changes, such as glycosylation, which is very similar to the human body. In the present study, firstly, the required primers were designed to amplify the gpdIIgene promoter. The genomic DNA was then extracted from mycelium of the button mushroom. After performing PCR with the pfupolymerase, a 709-bp fragment which contains gpdII promoter was replicated. In addition, the synthesized mini proinsulin gene (PInk) was transfered to E. coliand kept and recultured with bacteria itself. Both plasmids contained the components of the promoter of the gpdII(PgpdII) and the mini-pronsulin gene (PInk) and also the double digested pBGgHg plasmid and the bonding reaction wasperformed to link PgpdII with PInk and entering the resulted part to the pBGgHg plasmid. The recombinant plasmid was designated as pCAMB-hmPInk for confirmation of map structure in digestion enzyme

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